Instructions for Use

IGH + IGK B-Cell Clonality Assay
For identification of clonal immunoglobulin heavy chain and kappa light chain gene rearrangements.
For RESEARCH USE ONLY. Not for use in diagnostic procedures.

Manufactured in U.S.A.

Storage Conditions: -85ºC to -65ºC(DNA controls may be separated from assay kits and stored at 2°C to 8°C)

Catalog #11000010
11000031
11000041

Products

Quantity

IGH + IGK B-Cell Clonality Assay for Gel Detection
IGH + IGK B-Cell Clonality Assay for ABI Fluorescence Detection
IGH + IGK B-Cell Clonality Assay MegaKit for ABI Fluorescence Detection
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

33 Reactions
33 Reactions
330 Reactions

Page 2 of 21

Table of Contents
1.

ASSAY USE .........................................................................................................................................................................................32.

SUMMARY AND EXPLANATION OF THE TEST ...............................................................................................................................................3
2.1.
2.2.

3.

PRINCIPLES OF THE PROCEDURE ..............................................................................................................................................................3
3.1.
3.2.
3.3.

4.

Precautions ....................................................................................................................................................................................... 8Interfering Substances ..................................................................................................................................................................... 8Specimen Requirements and Handling ........................................................................................................................................... 8
Sample Preparation.......................................................................................................................................................................... 8
Sample Storage ................................................................................................................................................................................ 8
Template Amplification Control ...................................................................................................................................................... 8

ASSAY PROCEDURE ..............................................................................................................................................................................9
7.1.
7.2.
7.3.
7.4.
7.5.
7.6.
7.7.

8.

Thermal cycler .................................................................................................................................................................................. 7
Electrophoresis Unit......................................................................................................................................................................... 7
UV Illumination Unit......................................................................................................................................................................... 7
ABI Capillary Electrophoresis Instruments...................................................................................................................................... 7

SPECIMEN COLLECTION AND PREPARATION................................................................................................................................................8
6.1.
6.2.
6.3.
6.4.
6.5.
6.6.

7.

Reagent Components ...................................................................................................................................................................... 5
Warnings and Precautions ............................................................................................................................................................... 6
Storage and Handling ....................................................................................................................................................................... 7

INSTRUMENTS .....................................................................................................................................................................................7
5.1.
5.2.
5.3.
5.4.

6.

Polymerase Chain Reaction (PCR) ................................................................................................................................................... 3
Gel Electrophoresis .......................................................................................................................................................................... 4
Differential Fluorescence Detection ............................................................................................................................................... 4

REAGENTS..........................................................................................................................................................................................5
4.1.
4.2.
4.3.

5.

Background ....................................................................................................................................................................................... 3
Summary ........................................................................................................................................................................................... 3

Materials Provided ........................................................................................................................................................................... 9
Materials Required (not provided).................................................................................................................................................. 9Reagent Preparation ...................................................................................................................................................................... 11
Amplification .................................................................................................................................................................................. 11
Detection ........................................................................................................................................................................................ 11
Quality Control ............................................................................................................................................................................... 13
Recommended Positive Controls .................................................................................................................................................. 13

INTERPRETATION AND REPORTING .........................................................................................................................................................14
8.1.
8.2.

Sample Reporting ........................................................................................................................................................................... 14
Sample Interpretation .................................................................................................................................................................... 14

9.

LIMITATIONS OF PROCEDURE................................................................................................................................................................14

10.

EXPECTED VALUES .............................................................................................................................................................................15
10.1.
10.2.

Expected Size of Amplified Products............................................................................................................................................. 15
Sample Data ................................................................................................................................................................................... 16

11.

TECHNICAL AND CUSTOMER SERVICE......................................................................................................................................................19

12.

REFERENCES .....................................................................................................................................................................................19

13.

SYMBOLS .........................................................................................................................................................................................20

14.

LEGAL NOTICE ..................................................................................................................................................................................20
14.1.

15.

Warranty and Liability .................................................................................................................................................................... 20

IGH + IGK B-CELL CLONALITY ASSAY: SINGLE PAGE GUIDE ........................................................................................................................21

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 3 of 21

1. Assay Use
Immunoglobulin heavy chain (IGH) and Kappa light chain (IGK) gene rearrangement assays are useful for the study of identifying clonal
B-cell populations and evaluation of new research and methods in malignancy studies.

2. Summary and Explanation of the Test
2.1.

BackgroundRearrangements of the antigen receptor genes occur during ontogeny in B and T lymphocytes. These gene rearrangements
generate products that are unique in length and sequence for each cell. Therefore, polymerase chain reaction (PCR) assays canbe used to identify lymphocyte populations derived from a single cell by detecting the unique V-J gene rearrangements present
within these antigen receptor loci.1 This PCR assay employs multiple consensus DNA primers that target conserved genetic regions
within the immunoglobulin heavy chain gene. This test is used to detect the vast majority of clonal B-cell malignancies from DNA.
Test products can be analyzed using a variety of detection formats, including gel and capillary electrophoresis.
Invivoscribe’s Gene Rearrangement and Translocation Assays represent a simple approach to PCR-based clonality testing. Thesestandardized assays were carefully optimized testing positive and negative control samples using multiplex master mixes.

2.2.

SummaryThis test kit includes six (6) master mixes. IGH Tubes A, B, and C target the framework 1, 2, and 3 regions (respectively) within
the variable (VH) region and the joining (JH) region of the IGH locus. IGK Tubes A and B target the variable, intragenic and joining
regions of the IGK locus. The last master mix, the Specimen Control Size Ladder, targets multiple genes and generates a series ofamplicons of 96, 197, 297, 397, and 602 base pairs (bp) to ensure that the quality and quantity of input DNA is adequate to yielda valid result.
This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.

3. Principles of the Procedure
3.1.

Polymerase Chain Reaction (PCR)

Polymerase chain reaction (PCR) assays are routinely used for the identification of clonal B-cell populations. These tests amplify
the DNA between primers that target the conserved framework (FR) VH and JH regions (IGH Tubes A, B, and C) of the IGH locus,
the Vκ and Jκ regions (IGK Tube A) and the variable, intragenic and Kappa Deleting Element (Kde) regions (IGK Tube B) of the of
the IGK locus. These conserved regions lie on either side of an area within the V-J region where programmed genetic
rearrangements occur during maturation of all B and T lymphocytes.
The antigen receptor genes that undergo rearrangement are the immunoglobulin heavy chain and light chains genes in B-cells
and the T cell receptor genes in T-cells. Each B- and T-cell has a single productive V-J rearrangement that is unique in both lengthand sequence. Therefore, when this region is amplified using DNA primers that flank this region, a clonal population of cells yields
one or two prominent amplified products (amplicons) within the expected size range. Two products are generated in cases when
the initial rearrangement was non-productive and was followed by rearrangement of the other homologous chromosome. In
contrast, DNA from a normal or polyclonal (many clones) population produces a bell-shaped curve of amplicon products (Gaussian
distribution) that reflect the heterogeneous population of V-J region rearrangements.
Since the antigen receptor genes are polymorphic (consisting of a heterogeneous population of related DNA sequences), it isdifficult to employ a single set of DNA primer sequences to target all of the conserved flanking regions around the V-J
rearrangement. N-region diversity and somatic mutation further scramble the DNA sequences in these regions. Therefore
multiplex master mixes, which target several FR regions, are required to identify the majority of clonal rearrangements. As
indicated, clonal rearrangements are identified as prominent, single-sized products within the smear of different-sized amplicon
products that form a Gaussian distribution around a statistically favored, average-sized rearrangement. As expected, primers that

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 4 of 21

amplify from the different FR regions, which are located at three distinct regions along the heavy chain gene, produce a
correspondingly different size-range of V-J products.

Figure 1.

3.2.

Depicted is a simple representation of the organization of a rearranged immunoglobulin heavy chain (IGH) gene on
chromosome 14 and the immunoglobulin kappa light chain gene on chromosome 2p11.2. Black arrows represent the relative
positions of primers that target the conserved framework (FR1-3) and the downstream consensus JH gene segments for IGH
and the Vκ, Jκ, INTR and Kde primers which are included in the IGK master mix tubes. The amplicon products generated fromeach of these regions can be differentially detected when fluorescent primer sets are used with capillary electrophoresis
instruments that employ differential fluorescence detection.

Gel Electrophoresis

Gel electrophoresis, such as agarose gel electrophoresis or non-denaturing polyacrylamide gel electrophoresis (PAGE), is
commonly used to resolve the different amplicon products based on their size, charge, and conformation. Since DNA is negatively
charged, when an electrical potential (voltage) is applied across the gel containing PCR products, the electrical field causes theamplicons to migrate through the gel. Smaller DNA fragments are able to easily migrate through the gel matrix, whereas larger
DNA fragments migrate more slowly which causes a separation of the amplicon products based on size. Ethidium bromide orother DNA intercalating dyes can then be used to stain and detect these products in the gel.

3.3.

Differential Fluorescence Detection

Gel electrophoresis is commonly used to resolve the different-sized amplicon products and ethidium bromide or other DNA
intercalating dyes to stain and detect these products. A powerful alternative method is use of differential fluorescence detection
with primers conjugated with fluorescent dyes that correspond to different targeted regions. Reaction products from several
different master mixes can be pooled, fractionated using capillary electrophoresis and detected simultaneously. This detection
system results in unsurpassed sensitivity, single base resolution, differential product detection and relative quantification. In
addition, the laboratory can eliminate the use of agarose and polyacrylamide gels, as well as the use of carcinogens such as
ethidium bromide. Further, differential detection allows accurate, reproducible and objective interpretation of primer-specificproducts and automatic archiving of data. The automatic archiving of sample data allows comparison of data collected at different
times. The inter-assay and intra-assay reproducibility in size determination using capillary electrophoresis is approximately 1-2
base pairs.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 5 of 21

4. Reagents
4.1.

Reagent Components

Table 1: Available Kits

Description

Catalog #Quantity

11000010

IGH Gene Clonality Assay – Gel Detection

33 Reactions

11000031

IGH Gene Clonality Assay – ABI Fluorescence Detection

33 Reactions

11000041

IGH Gene Clonality Assay MegaKit – ABI Fluorescence Detection

330 Reactions

Table 2: Kit Components

Reagent

Catalog #40881750
Positive Control
DNAs

40881090
40880370

Negative (Normal)
Control DNA

40920010

Reagent Components(active ingredients)
IVS-0030 Clonal Control DNA

200 µg/mL of DNA in 1/10th TE solution

IVS-0019 Clonal Control DNA

200 µg/mL of DNA in 1/10th TE solution

IVS-0007 Clonal Control DNA

200 µg/mL of DNA in 1/10th TE solution

IVS-0000 Polyclonal Control DNA

200 µg/mL of DNA in 1/10th TE solution

Unit
Quantity

Assay Kit
# of Units

Assay
MegaKit# of Units

100 µL1

5100 µL

1

5

StorageTemp.

or100 µL1

5100 µL1

51500 µL

1

N/A

1500 µL

1

N/A

1500 µL

1

N/A

1500 µL

1

N/A

1500 µL

1

N/A

1500 µL

1

N/A

Gel Detection
IGH Tube A – Unlabeled21010010

Multiple oligonucleotides targeting theframework 1 region of the immunoglobulin heavy
chain gene in a buffered salt solution.

IGH Tube B – Unlabeled21010020

Multiple oligonucleotides targeting theframework 2 region of the immunoglobulin heavy
chain gene in a buffered salt solution.

IGH Tube C – Unlabeled
Master Mixes forGel Detection

21010030

Multiple oligonucleotides targeting theframework 3 region of the immunoglobulin heavy
chain gene in a buffered salt solution.

IGK Tube A - Unlabeled21020010

Multiple oligonucleotides targeting the variable
and joining regions of the immunoglobulin kappalight chain gene in a buffered salt solution.

IGK Tube B - Unlabeled21020020

Template
Amplification
Control Master Mix

20960020

Multiple oligonucleotides targeting the variable,
Jκ-Cκ intron, and Kde regions of theimmunoglobulin kappa light chain gene in abuffered salt solution.

Specimen Control Size Ladder – Unlabeled

Multiple oligonucleotides targeting housekeeping
genes.

Differential Fluorescence Detection
IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 6 of 21
Table 2: Kit Components

Reagent

Reagent Components(active ingredients)

Catalog #Unit
Quantity

Assay Kit
# of Units

Assay
MegaKit# of Units

1500 µL

1

101500 µL

1

101500 µL

1

101500 µL

1

101500 µL

1

101500 µL

1

10

Storage
Temp.

IGH Tube A – 6FAM
21010011

Multiple oligonucleotides targeting theframework 1 region of the immunoglobulin
heavy chain gene in a buffered salt solution.

IGH Tube B – 6FAM
21010101

Master Mixes forDifferential
Fluorescence
Detection

Multiple oligonucleotides targeting theframework 2 region of the immunoglobulin
heavy chain gene in a buffered salt solution.

IGH Tube C – HEX
21010031

Multiple oligonucleotides targeting theframework 3 region of the immunoglobulin
heavy chain gene in a buffered salt solution.

IGK Tube A - 6FAM
21020011

Multiple oligonucleotides targeting the variable
and joining regions of the immunoglobulin kappalight chain gene in a buffered salt solution.

IGK Tube B - 6FAM

21020021

Template
Amplification
Control Master Mix

4.2.

20960021

Multiple oligonucleotides targeting the variable,
Jκ-Cκ intron, and Kde regions of theimmunoglobulin kappa light chain gene in abuffered salt solution.

Specimen Control Size Ladder – 6FAM

Multiple oligonucleotides targeting housekeeping
genes.

Warnings and Precautions

















For RESEARCH USE ONLY. Not for use in diagnostic procedures.
Use this assay kit as a system. Do not substitute other manufacturer’s reagents. Dilution, reducing amplification
reaction volumes, or other deviation in this protocol may affect the performance of this test and/or nullify any
limited sublicense that comes with the purchase of this testing kit.
Materials are stable until the labeled expiration date when stored and handled as directed. Do not use kits beyond
their expiration date.
Close adherence to the protocol will assure optimal performance and reproducibility. Use care to ensure use ofcorrect thermal cycler program, as other programs may provide inaccurate/faulty data, such as false positive andfalse negative results.
Do not mix or combine reagents from kits with different lot numbers.
Wear appropriate personal protective equipment and follow good laboratory practices and universal precautions
when working with specimens. Handle specimens in approved biological safety containment facilities and open onlyin certified biological safety cabinets. Use molecular biology grade water for the preparation of specimen DNA.
Due to the analytical sensitivity of this test, use extreme care to avoid the contamination of reagents or amplification
mixtures with samples, controls or amplified materials. Closely monitor all reagents for signs of contamination (e.g.,
negative controls giving positive signals). Discard reagents suspected of contamination.
To minimize contamination, wear clean gloves when handling samples and reagents and routinely clean work areasand pipettes prior to doing PCR.
Autoclaving does not eliminate DNA contamination. Follow uni-directional workflow in the PCR laboratory; begin
with master mix preparation, move to specimen preparation, then to amplification, and finally to detection. Do notbring amplified DNA into the areas designated for master mix or specimen preparation.
Dedicate all pipettes, pipette tips, and any equipment used in a particular area to that area of the laboratory.
Use sterile, disposable plastic ware whenever possible to avoid RNase, DNase, or cross-contamination.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 7 of 21

4.3.

Storage and Handling






For any duration other than immediate use, store assay kits at -85°C to -65°C.
The optimum storage temperature for DNA controls is 2°C to 8°C, but DNA controls can be stored at -85°C to -65°C
long term.
All reagents and controls must be thawed and vortexed or mixed thoroughly prior to use to ensure that they areresuspended completely. Excessive vortexing may shear DNA and cause labeled primers to lose their fluorophors.
Materials are stable until the labeled expiration date when stored and handled as directed. Do not use kits beyond
their expiration date.
Due to high salt concentrations, PCR master mixes are sensitive to freeze/thaw cycles. Aliquot master mixes into
sterile o-ring screw-cap tubes if necessary.

5. Instruments
5.1.

Thermal cyclerUse or function: Amplification of DNA samples
Suggested instrument: Veriti™ Thermal Cycler or equivalent
Performance characteristics and specification:
o
o




5.2.

Follow manufacturer’s installation, operation, calibration, and maintenance procedures.
See section 7.4 Amplification for thermal cycler program.

Electrophoresis Unit

Use or function: DNA fragment separation
Performance characteristics and specification:
o



5.3.

Capable of running at 35V to 135V for extended times

Follow manufacturer’s installation, operation, calibration, and maintenance procedures.

UV Illumination Unit

Use or function: DNA detection
Performance characteristics and specification:
o



5.4.

Minimum thermal range: 15°C to 96°C
Minimum ramping speed: 0.8°C/sec

Capable of emitting light at a wavelength of ~302 nm

Follow manufacturer’s installation, operation, calibration, and maintenance procedures.

ABI Capillary Electrophoresis Instruments

Use or function: Fragment detection and analysis
Performance characteristics and specification:o

The following capillary electrophoresis instruments will meet the performance needs for this assay:
•
•
•
•
•
•
•






ABI 310 Genetic Analyzer (1-capillary)
ABI 3100 Avant Genetic Analyzer (4-capillaries)
ABI 3100 Genetic Analyzer (16-capillaries)
ABI 3130 Genetic Analyzer (4-capillaries)
ABI 3130xL Genetic Analyzer (16-capillaries)
ABI 3500 Genetic Analyzer (8-capillaries)
ABI 3500xL Genetic Analyzer (24-capillaries)

Follow manufacturer’s installation, operation, calibration, and maintenance procedures.
The ABI instrument used must be calibrated with appropriate Matrix Standards as outlined in section 7.2 Materials
Required (not provided).
Use the default settings for your polymer and capillary type.
See section 7.5 Detection.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 8 of 21

6. Specimen Collection and Preparation
6.1.

Precautions

Biological specimens from humans may contain potentially infectious materials. Handle all specimens in accordance with the
OSHA Standard on Bloodborne Pathogens or Biosafety Level 2.

6.2.

Interfering Substances

The following substances are known to interfere with PCR:





6.3.

Divalent cation chelators
Low retention pipette tips
EDTA (not significant at low concentrations)
HeparinSpecimen Requirements and HandlingThis assay tests extracted and purified genomic DNA (gDNA). Common sources of gDNA include:



6.4.

5 cc of peripheral blood, bone marrow biopsy or bone marrow aspirate anti-coagulated with heparin or EDTA. Shipat ambient temperature; OR
Formalin-fixed paraffin embedded tissue or slides.

Sample Preparation
6.4.1.

Using any method of DNA extraction, extract the gDNA from unknown samples.

6.4.2.

Resuspend DNA to final concentration of 100 μg – 400 μg per mL in TE (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) or
distilled water.



6.5.

This is a robust assay system. A wide range of DNA concentrations will generate a valid result. Therefore,
quantifying and adjusting DNA concentrations is generally not necessary.
When testing unknown samples, a template amplification control master mix (e.g., Specimen Control Sizeladder) can be used to verify the absence of PCR inhibitors and sufficient quality and quantity of DNA ispresent to generate a valid result.

Sample Storage

Store samples using a method that prevents degradation of DNA.

6.6.

Template Amplification Control
6.6.1.

The Specimen Control Size Ladder master mix primers are available unlabeled for gel detection and labeled with afluorescent dye (6-FAM) for ABI fluorescence detection. The label is detected as BLUE using the differential
fluorescence software. The amplicons produced with this master mix are at 96, 197, 297, 397, and 602 base pair
(bp).



The ~96 bp band is comprised of 84 bp and 96 bp bands. Both of these bands co-migrate on a gel.
Run the products of this master mix separately.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 9 of 21

7. Assay Procedure
7.1.

Materials Provided

Table 3: Kit components

IGH + IGK B-Cell Clonality Assay for ABI Fluorescence Detection

IGH + IGK B-Cell Clonality Assay for Gel Detection
Catalog #Description

Catalog #Description

21010010

IGH Tube A – Unlabeled21010011

IGH Tube A – 6FAM

21010020

IGH Tube B – Unlabeled21010101

IGH Tube B – 6FAM

21010030

IGH Tube C – Unlabeled21010031

IGH Tube C – HEX

21020010

IGK Tube A – Unlabeled21020011

IGK Tube A – 6FAM

21020020

IGK Tube B – Unlabeled21020021

IGK Tube B – 6FAM

20960020

Specimen Control Size Ladder – Unlabeled20960021

Specimen Control Size Ladder – 6FAM

40881750

IVS-0030 Clonal Control DNA

40881750

IVS-0030 Clonal Control DNA

40881090

IVS-0019 Clonal Control DNA

40881090

IVS-0019 Clonal Control DNA

40880370

IVS-0007 Clonal Control DNA

40880370

IVS-0007 Clonal Control DNA

40920010

IVS-0000 Polyclonal Control DNA

40920010

IVS-0000 Polyclonal Control DNA

7.2.

Materials Required (not provided)

Table 4: Materials Required (not provided)

Reagent/Material

DNA Polymerase

Recommended Reagents/Materials and Suppliers
Roche:• EagleTaq DNA Polymerase
Invivoscribe:• FalconTaq DNA Polymeraseor equivalent

Glass Distilled De-ionized
Molecular Biology Gradeor USP Water

Calibrated Pipettes

Catalog Number

Notes

05206944190
60970130

N/A

Rainin:• P-2, P-20, P-200, and P-1000 pipettes• Or SL-2, SL-20, SL-200, and SL-1000 pipettes

N/A

N/A

Sterile and free of DNase andRNase.

N/A

Must be able to accurately
measure volumes between 1 µL
and 1000 µL.

N/A

N/A

Thermo Fisher Scientific:• Veriti Dx Thermal Cycler
Thermal cycler

Bio-Rad:• MJ Research PTC-100 or PTC-200, PTC-220, PTC-240
Perkin-Elmer• PE 9600 or PE 9700

Vortex Mixer

N/A

N/A

N/A

PCR plates or tubes

N/A

N/A

SterileFilter barrier pipette tips

N/A

N/A

Sterile, RNase/DNase/Pyrogenfree

Microcentrifuge tubes

N/A

N/A

SterileGel Detection

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 10 of 21
Table 4: Materials Required (not provided)

Reagent/Material

Recommended Reagents/Materials and Suppliers

Catalog Number

Notes

Gel Electrophoresis Unit

N/A

N/A

For polyacrylamide gels

Ethidium Bromide
6% Polyacrylamide Gels

Thermo Fisher Scientific:• UltraPureTM 10 mg/mL Ethidium Bromide

5585-011

Thermo Fisher Scientific:•

Novex TBE Gels (6%, 12 well)EC62652Box

TBE Running Buffer

Thermo Fisher Scientific:• Novex TBE Running Buffer (5X)

LC6675

Gel Loading Buffer

Thermo Fisher Scientific:• 10X BlueJuiceTM Gel Loading Buffer• Novex Hi-Density TBE Sample Buffer (5X)

10816-015
LC6678100 bp DNA Ladder

Thermo Fisher Scientific:• TrackItTM 100 bp DNA Ladder

10488-058

N/A
N/A
Dilute 1:5 prior to use.

N/A

N/A

Differential Fluorescence Detection
ABI Capillary
Electrophoresis
Instrument

Thermo Fisher Scientific:• ABI 310, 3100, or 3500 series

Hi-Di Formamide

Thermo Fisher Scientific:• Hi-DiTM Formamide

4311320

Invivoscribe:• Hi-Di Formamide w/ROX size standards for ABI 310• Hi-Di Formamide w/ROX size standards for ABI 3100

60980051
60980061

Size Standards

Thermo Fisher Scientific:• For ABI 3100 or 3130 instruments:
o GeneScanTM - 400HD [ROX]TM•For ABI 3500 instruments:
o GeneScan - 600 [LIZ]TM v2.0

Thermo Fisher Scientific:• For ABI 3100 and 3130 instruments:o DS-30 Matrix Standard Kit (Dye Set D)•

Spectral Calibration Dye
Sets•

Polymer

44083994345827
402996401546

Thermo Fisher Scientific:• POP-4TM Polymer:o POP-4 for 310 Genetic Analyzerso POP-4 for 3100/3100-Avant Genetic Analyzerso POP-4 for 3130/3130xL Genetic Analyzers

40283843163554352755

Thermo Fisher Scientific:• 10X Genetic Analyzer Buffer with EDTA

N/A

4029854345833

POP-7TM Polymer:o POP-7 for 3130/3130xL Genetic Analyzerso POP-7 for 3500/3500xL Genetic Analyzers

N/A

N/A

For ABI 3500 instruments:o DS-33 Matrix Standard Kit (Dye Set G5)•

BufferFor ABI 310 instruments:
o NED Matrix Standardo And Fluorescent Amidite Matrix Standards [6FAM, TET,HEX, TAMRA, ROX]

N/A

Dye set used to spectrally calibrate
ABI instrument for use with 6FAM,
HEX, NED, and ROX

N/A

43527594393714402824

IGH + IGK B-Cell Clonality Assay

Dilute 1:10 in sterile water beforeuse280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 11 of 21

7.3.

Reagent PreparationAll unknown samples can be tested using the Specimen Control Size ladder to ensure that no inhibitors of
amplification are present and there is DNA of sufficient quality and quantity to generate a valid result.
Test all samples in singlicate.
Test positive, negative and no template controls for each master mix.
Batch multiple samples in a run to avoid running out of the negative control (IVS-0000 Polyclonal Control DNA). If
batching samples is not practical, IVS-0000 Polyclonal Control DNA is also available for purchase separately.

7.3.1.

Using gloved hands, remove the master mixes from the freezer. Allow the tubes to thaw; then gently vortex tomix.

7.3.2.

In containment hood or dead air box, remove an appropriate aliquot to clean, sterile microfuge tube (one tube
for each master mix).


7.3.3.

Add the appropriate amount of Taq DNA polymerase (0.25 μL of @5 U/μL per 50 μL total reaction volume) toeach master mix and gently mix by inverting several times or gently vortexing.


7.4.

Aliquot volumes = 45 μL for each sample + 135 μL (3 x 45 μL) for the positive, negative and no templatecontrols.
Include an additional 20 μL to correct for pipetting errors.

The master mixes are now ready for distribution to reaction tubes or plate and addition of sample.

Amplification
7.4.1.

Aliquot 45 μL of the master mix/enzyme solutions into individual PCR wells or tubes.

7.4.2.

Add 5 μL of sample or control DNA to the individual tubes or wells containing the respective master mix reactions.


7.4.3.

Pipette up and down several times to mix.

Amplify the reactions using the following PCR program.
Use the calculated option for temperature measurement with the PTC instruments.
Table 5: Thermal cycling conditions

7.4.4.

7.5.

Step

Temperature

Duration

Cycles

1
2
3
4
5
6

95°C
95°C
60°C
72°C
72°C
15°C

7 minutes
45 seconds
45 seconds
90 seconds
10 minutes
∞

1
35
1
1

Remove the amplification plate from the thermal cycler.

Detection


Not all detection formats are available for all assays.

Gel Detection – Polyacrylamide TBE Gels
7.5.1.

Assemble electrophoresis unit using a 6% non-denaturing polyacrylamide TBE gel (made with 1X TBE) and 0.5X
TBE running buffer.

7.5.2.

Add 5 μL of ice-cold non-denaturing bromophenol blue loading buffer to samples.

7.5.3.

Load 20 μL of mixture into wells of the gel.

7.5.4.

Run gel at 110V for 2-3 hours or 40-50V overnight.
Voltage and electrophoresis time depend on the PCR amplicon size, acrylamide gel thickness and type of PCR
equipment.
Voltage and run time can be adapted accordingly.

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7.5.5.

Stain gels in 0.5 µg/mL ethidium bromide (in water or 0.5X TBE Buffer) for 5-10 minutes.

7.5.6.

Destain gels 2X in water for 5-10 minutes.

7.5.7.

Visualize the gel using UV illumination.

7.5.8.

Photograph the gel and interpret the data.

Gel Detection – Heteroduplex Analysis (RECOMMENDED)

Do not heteroduplex the PCR products from the Specimen Control Size Ladder master mix. Skip steps 7.5.9 - 7.5.10
and proceed with step 7.5.11.

7.5.9.

Denature 20 μL of PCR products at 94ºC for 5 minutes.

7.5.10.

Re-anneal PCR products at 4ºC for 60 minutes.

7.5.11.

Assemble electrophoresis unit using a 6% non-denaturing polyacrylamide TBE gel (made with 1X TBE) and 0.5X
TBE running buffer.

7.5.12.

Add 5 μL of ice-cold non-denaturing bromophenol blue loading buffer to samples

7.5.13.

Load 20 μL of mixture into wells of the gel.

7.5.14.

Run gel at 110V for 2-3 hours or 40-50V overnight.
Voltage and electrophoresis time depend on the PCR amplicon size, acrylamide gel thickness and type of PCR
equipment.
Voltage and run time can be adapted accordingly.

7.5.15.

Stain gels in 0.5 µg/mL ethidium bromide (in water or 0.5X TBE Buffer) for 5-10 minutes.

7.5.16.

Destain gels 2X in water for 5-10 minutes.

7.5.17.

Visualize the gel using UV illumination.

7.5.18.

Photograph the gel and interpret the data.

ABI Fluorescence Detection with ABI 310, 3100 & 3130 instruments
7.5.19.

In a new microcentrifuge tube, mix an appropriate amount (10 μL per PCR reaction) of Hi-Di Formamide with ROX
Size Standards. Vortex well.

7.5.20.

In a new 96-well PCR plate, add 10 μL of Hi-Di Formamide with ROX size standards to individual wells for eachreaction.

7.5.21.

Transfer 1 μL of each reaction to the wells containing Hi-Di Formamide with ROX size standards.


Add only one sample per well.
Pipette up and down to mix.

7.5.22.

Cap or cover the PCR plate.

7.5.23.

Heat denature the samples at 95ºC for 2 minutes, then snap chill on ice for 5 minutes.

7.5.24.

Prepare a sample sheet and injection list for the samples.

7.5.25.

Run the samples on an ABI 310/3100/3130 capillary electrophoresis instrument according to its user manual.


7.5.26.

Data are automatically displayed as size and color specific peaks.

Review profile and controls, report results.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 13 of 21

ABI Fluorescence Detection with ABI 3500 instruments

7.6.

Note:

Due to instrument to instrument variation in the performance of the ABI 3500 platform, the amount of formamide,
sample and size standard listed in the protocol is intended to be a starting point. The protocol may need to be
optimized for specific ABI 3500 Platforms.

7.5.27.

In a new microcentrifuge tube, mix an appropriate amount (9.5 μL per PCR reaction) of Hi-Di Formamide with LIZ
Size Standards. Vortex well.

7.5.28.

In a new 96-well PCR plate, add 9.5 μL of Hi-Di Formamide with LIZ size standards to individual wells for each PCR
reaction.

7.5.29.

Transfer 0.5 μL of each PCR reaction to the wells containing Hi-Di Formamide with LIZ size standards. Add only
one sample per well. Pipette up and down to mix.

7.5.30.

Cap or cover the PCR plate.

7.5.31.

Heat denature the samples at 95ºC for 3 minutes, then snap chill on ice for 5 minutes.

7.5.32.

Prepare a sample sheet and injection list for the samples.

7.5.33.

Run the samples on an ABI 3500 capillary electrophoresis instrument according to its user manual.

7.5.34.

Data are automatically displayed as size and color specific peaks. Review profile and controls, report results.

Quality Control

Positive and negative (or normal) controls are furnished with the kit and can be run in singlicate each time the assay is performed
to ensure proper performance of the assay. Include a no template control (e.g. water) to test for contamination of the mastermix or cross-contamination of PCR. A buffer control may also be added to ensure that no contamination of the buffer used toresuspend the samples has occurred. The values for the positive controls are provided under section 10.1 Expected Size of
Amplified Products. Additional controls and sensitivity controls (dilutions of positive controls into our negative control) areavailable from Invivoscribe, Inc.

7.7.

Recommended Positive Controls

Table 6: Recommended Positive Controls

Master Mix

TargetColorControl DNA

Catalog #Product Size in (bp)

IGH Tube A

FR1-JHBlue

Valid Size Range
IVS-0030 Clonal Control DNA--40881750

310 - 360
2801, 325

IGH Tube B

FR2-JHBlue

Valid Size Range
IVS-0030 Clonal Control DNA--40881750

250 - 2952
260

IGH Tube C

FR3-JHGreen

Valid Size Range
IVS-0019 Clonal Control DNA--40881090

100 - 170
145

IGK Tube A

Vκ-Jκ

Blue

Valid Size Range
IVS-0007 Clonal Control DNA--40880370

120 - 160, 190 - 210, 260 - 300
143

IGK Tube B

Vκ-Kde +intron-Kde

Blue

Valid Size Range
IVS-0007 Clonal Control DNA--40880370

210 - 250, 270 - 300, 350 - 390
274, 282

Specimen Control Size
LadderMultiple
Genes

Blue

Valid Size Range
IVS-0000 Polyclonal Control DNA--40920010

84, 96, 197, 297, 397, 6023
84, 96, 197, 297, 397, 602

Note:
1Note:
2Note:
3Note:

The amplicon sizes listed above were determined using an ABI platform. Amplicon sizes seen on your specific CE instrument may differ 1-4 bp fromthose listed above depending on the platform of detection (ABI) and the version of the analysis software used. Once identified, the amplicon size as
determined on your specific platform will be consistent from run to run.
A 280 bp band may also be present and is a known amplicon that lies just outside the valid size range for IGH Tube A.
A non-specific peak may be observed at 227 bp.
Because smaller PCR fragments are preferentially amplified, it is not unusual for the 602 bp fragment to have a diminished signal or to be missing
entirely.

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8. Interpretation and Reporting
This assay is for research use only. PCR based testing does not identify 100% of clonal cell populations; therefore, repeat testing by
Next Generation Sequencing may be advisable to rule out clonality.

8.1.

Sample Reporting

Results can be reported as “Positive” or “Negative” for “Detection of clonal immunoglobulin heavy chain or kappa light chain generearrangement consistent with the presence of a clonal cell population”.
8.1.1.

Report samples that fail to amplify following repeat testing as “A result cannot be reported on this specimen
because there was DNA of insufficient quantity or quality for analysis”.

8.1.2.

All assay controls must be examined prior to interpretation of sample results. If the controls do not yield thecorrect results, the assay is not valid and the samples cannot be interpreted.

The following describes the analysis of each of the controls and the decisions necessary based upon the results.


Negative Control:(Polyclonal control, water or no template blank). If the negative control is:oPositive:o

Negative:

Possible contamination of all PCR amplification reactions. Do not continue with theinterpretation of results. Prepare fresh master mix and repeat amplification.
Continue with the analysis.

Positive Control:(This can also be an extraction control if positive control material is taken through extraction
processes). If the positive control is:ooContinue with analysis.
Repeat the assay.

Specimen Control Size Ladder: (This is run on unknown samples only). If the amplification control:
o
o

8.2.

Positive:
Negative:

Generates ~96, 197, 297, 397, and 602 bp products:
Because smaller PCR fragments are preferentially
amplified, it is not unusual for the 602 bp fragment to have a diminished signal or to be missing
entirely. Continue with analysis.
Does not generate specified products:
Repeat assay and/or re-extract the specimen.

Sample Interpretation

Following the acceptance of the controls, the samples are interpreted as follows:
8.2.1.

One or two prominent bands within the valid size range for IGH Tubes A, B, or C is reported as “Detection of clonal
immunoglobulin heavy chain gene rearrangement consistent with the presence of a clonal cell population”.

8.2.2.

One or two prominent bands within the valid size range for IGK Tubes A or B is reported as “Detection of clonal
immunoglobulin kappa light chain gene rearrangement consistent with the presence of a clonal cell population”.

9. Limitations of Procedure
The assay is subject to interference by degradation of DNA or inhibition of PCR due to heparin or other agents.

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10. Expected Values
10.1.

Expected Size of Amplified Products

The size range for each of the master mixes has been determined testing positive control samples. For accurate and meaningfulinterpretation it is important to ignore peaks that occur outside of the proscribed/valid size range for each of the master mixes.
Peaks that are outside of the range cannot be assumed to be valid.
Note:“Color” indicates the color of products generated with the master mix when using differential fluorescence detection
format (e.g., ABI instruments).

Table 7: Expected size of Amplified Products

Master Mix

TargetColorControl DNA

Catalog
NumberProduct Size (bp)

IGH Tube A

FR1-JHBlue

Valid Size Range
IVS-0000 Polyclonal Control DNA
IVS-0030 Clonal Control DNA
IVS-0019 Clonal Control DNA--40920010
40881750
40881090

310 - 36085, 310 - 360
2801, 325
345

Blue

Valid Size Range
IVS-0000 Polyclonal Control DNA
IVS-0030 Clonal Control DNA
IVS-0019 Clonal Control DNA--40920010
40881750
40881090

250 - 2952
250 - 295260
285--40920010
40881750
40881090

100 - 170
100 - 170--145

IGH Tube B

FR2-JH

IGH Tube C

FR3-JHGreen

Valid Size Range
IVS-0000 Polyclonal Control DNA
IVS-0030 Clonal Control DNA
IVS-0019 Clonal Control DNA

IGK Tube A

Vκ-Jκ

Blue

Valid Size Range
IVS-0000 Polyclonal Control DNA
IVS-0007 Clonal Control DNA--40920010
40880370

120 - 160, 190 - 210, 260 - 300
135 - 155
143

IGK Tube B

Vκ-KdeBlue

Valid Size Range
IVS-0000 Polyclonal Control DNA
IVS-0007 Clonal Control DNA--40920010
40880370

210 - 250, 270 - 300, 350 - 390
225 - 245, 265 - 285, 4043
274, 282

Specimen Control
Size LadderMultiple
Genes

Blue

Any Human DNA

Note:
1Note:

Note:
Note:

2
3

---

84, 96, 197, 297, 397, 602

The amplicon sizes listed above were determined using an ABI platform. Amplicon sizes seen on your specific CE instrument may differ 1-4 bp fromthose listed above depending on the platform of detection and the version of the analysis software used. Once identified, the amplicon size as
determined on your specific platform will be consistent from run to run.
The 280 bp band is a known amplicon that lies just outside the valid size range for Tube A.
A non-specific peak may be observed at 227 bp.
Under sub-optimal conditions a nonspecific product of 404 bp can be detected in Tube B. To discriminate between specific and nonspecific,
negative control DNA will not show this band within the same experiment. If a band is present, we then consider the band nonspecific.

IGH + IGK B-Cell Clonality Assay

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10.2.

Sample Data

Gel Detection
The data shown in Figures 2 -7 were generated using the master mixes indicated:

Figure 2.

The data shown above was generated using
the IGH Tube A master mix and run on a 6%
polyacrylamide gel.

Figure 3.

The data shown above was generatedusing the IGH Tube B master mix and run
on a 6% polyacrylamide gel.

Figure 4.

The data shown above was generated using the
IGH Tube C master mix and run on a 6%
polyacrylamide gel.

Figure 5.

The data shown above was generated using
the IGK Tube A master mix and run on a 6%
polyacrylamide gel.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 17 of 21

Figure 6.

The data shown above was generated using the
IGK Tube B master mix and run on a 6%
polyacrylamide gel.

Figure 7.

The data shown above was generated using theSpecimen Size Control Ladder master mix and run on a
6% polyacrylamide gel.

ABI Fluorescence Detection
The data shown in Figures 8 - 12 were run on an ABI instrument using the master mixes indicated..

Figure 8.

Figure 9.

The data shown above was generated using
the IGH Tube A – 6FAM master mix.

The data shown above was generated using the
IGH Tube B– 6FAM master mix.

IGH + IGK B-Cell Clonality Assay

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Figure 11.

Figure 10.

The data shown above was generated using
the IGH Tube C– HEX master mix.

Figure 12.

The data shown above was generated using
the IGK Tube B– 6FAM master mix.

The data shown above was generated using the
IGK Tube A– 6FAM master mix.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 19 of 21

Figure 13.

11.

The data shown above was generated using theSpecimen Control Size Ladder– 6FAM mastermix.

Technical and Customer Service

Thank you for purchasing our IGH + IGK B-Cell Clonality Assay. We appreciate your business. We are happy to assist you in thevalidation of this assay and will provide ongoing technical assistance to keep the assays performing efficiently in your laboratory.
Technical assistance is most rapidly obtained using our Internet site: http://www.invivoscribe.com or by sending an email inquiry to:support@invivoscribe.com.
Contact Information
Invivoscribe, Inc
10222 Barnes Canyon Road | Building 1 | San Diego | California 92121-2711 | USA
Phone: +1 858 224-6600 | Fax: +1 858 224-6601 | Business Hours: 7:00AM - 5:00 PM PST/PDT
Technical Service: support@invivoscribe.com | Customer Service: sales@invivoscribe.com | Website: www.invivoscribe.com

12. References
1.

Rock, E.P., Sibbald, P.R., Davis, M.M. and Chien, Y. (1994). CDR3 Length in Antigen-specific Immune Receptors. J Exp Med.
179, 323-328
1. Miller, JE, Wilson, SS, Jaye, DJ, Kronenberg, M. (1999). An automated semiquantitative B and T cell clonality assay. MolecularDiagnostics 4, 101-117.
2. Van Dongen, JJM et al. (2003). Design and standardization of PCR primers and protocols for detection of clonal
immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: Report of the BIOMED2 Concerted Action BMH4-CT98-3936. Leukemia. 17, 2257-2317.
2. van Krieken, JHJM et al. (2007). Improved reliability of lymphoma diagnostics via PCR-based clonality testing: – Report ofthe BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia 21, 201-206.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures

Page 20 of 21

13. Symbols
The following symbols are used in labeling for Invivoscribe products.
Catalog Number

Expiration Date

Reagent Volume

Manufacturer

Lot Number

Consult Instructions for Use

Storage Conditions

Research Use Only

14. Legal Notice
14.1.

Warranty and Liability

Invivoscribe, Inc. (Invivoscribe®) is committed to providing the highest quality products.Invivoscribe® warrants that the products
meet or exceed the performance standards described in the Instructions For Use, as to products with such an insert. If a productis covered by product specifications and does not perform as specified, our policy is to replace the product or credit the fullpurchase price. No other warranties of any kind, expressed or implied, are provided by Invivoscribe®. Invivoscribe® liability shallnot exceed the purchase price of the product. Invivoscribe shall have no liability for direct, indirect, consequential or incidental
damages arising from the use, results of use, or inability to use its products; product efficacy under purchaser controlled
conditions in purchaser’s laboratory must be established and continually monitored through purchaser defined and controlled
processes including but not limited to testing of positive, negative, and blank controls every time a sample is tested. Ordering,
acceptance, and use of product constitutes purchaser acceptance of sole responsibility for assuring product efficacy andpurchaser agreement to the limitation of liability set forth in this paragraph.
This product is for Research Use Only; not for use in diagnostic procedures.
This product is covered by one or more of the following: European Patent Number 1549764, European Patent Number 2418287,
European Patent Number 2460889, Japanese Patent Number 4708029, United States Patent 8859748, and related pending andfuture applications. All of these patents and applications are licensed exclusively to Invivoscribe®. Additional patents licensed to
Invivoscribe covered in some of these products apply elsewhere. Many of these products require nucleic acid amplification
methods such as Polymerase Chain Reaction (PCR). No license under these patents to use amplification processes or enzymes isconveyed expressly or by implication to the purchaser by the purchase of this product.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
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Page 21 of 21

15. IGH + IGK B-Cell Clonality Assay: Single Page Guide
15.1.
15.2.

Using gloved hands, remove the master mixes from the freezer. Allow the tubes to thaw; then gently vortex to mix.
In a containment hood or dead air box remove an appropriate aliquot to clean, sterile microfuge tube (one tube for each of
the master mixes).



15.3.
15.4.
15.5.
15.6.

Aliquot volumes = 45 μL for each sample + 135 μL for the positive, negative and no template controls.
Add an additional 20 μL to correct for pipetting errors.

Add the appropriate amount of Taq DNA polymerase (0.25 μL of Taq @5 U/μL per 50 μL total reaction volume) to each mastermix and gently mix by inverting several times or gently vortexing.
Aliquot 45 μL of master mix to individual wells of a PCR plate.
Add 5 μL of DNA from the unknown and control samples to individual tubes or wells containing the respective master mix
reactions and pipette up and down several times to mix.
Amplify target DNA using the universal thermal cycler program.

Gel Detection – Heteroduplex Analysis
15.7.
15.8.
15.9.
15.10.
15.11.
15.12.
15.13.
15.13.
15.14.
15.15.

Denature 20 μL PCR products at 94ºC for 5 minutes.
Re-anneal PCR products at 4ºC for 60 minutes.
Assemble electrophoresis unit using a 6% non-denaturing polyacrylamide TBE gel (made with 1X TBE) and 0.5X TBE running buffer.
Add 5 μL ice-cold non-denaturing bromophenol blue loading buffer to samples
Load 20 μL mixture into wells of the gel.
Run gel at 110V for 2-3 hours or 40-50V overnight.
Stain gels in 0.5 µg/mL ethidium bromide (in water or 0.5X TBE Buffer) for 5-10 minutes.
Destain gels 2X in water for 5-10 minutes.
Use UV illumination to visualize gel.
Photograph the gel and interpret the data.

ABI Fluorescence Detection with ABI 310, 3100 & 3130 instruments
15.16.
15.17.
15.18.

In a new microcentrifuge tube, mix an appropriate amount (10 μL per reaction) of Hi-Di Formamide with ROX Size Standards.
Vortex well.
In a new 96-well PCR plate, add 10 μL Hi-Di Formamide with ROX size standards to individual wells for each reaction.
Transfer 1 μL of each reaction to the wells containing Hi-Di Formamide with ROX size standards.


15.19.
15.20.
15.21.
15.22.
15.23.

Add only one sample per well.
Pipette up and down to mix.

Cap or cover the PCR plate.
Heat denature the samples at 95 ºC for 2 minutes, then snap chill on ice for 5 minutes.
Prepare a sample sheet and injection list for the samples.
Run the samples on an ABI 310/3100/3130 capillary electrophoresis instrument according to its user manual.
Review profile and controls, report results.

ABI Fluorescence Detection with ABI 3500 instruments
Note: Due to instrument to instrument variation in the performance of the ABI 3500 platform, the amount of formamide, sample and size
standard listed in the protocol is intended to be a starting point. The protocol may need to be optimized for specific ABI 3500
Platforms.
15.24.
15.25.
15.26.

In a new microcentrifuge tube, mix an appropriate amount (9.5 μL per reaction) Hi-Di Formamide with LIZ Size Standards.
Vortex well.
In a new 96-well PCR plate, add 9.5 μL Hi-Di Formamide with LIZ size standards to individual wells for each reaction.
Transfer 0.5 μL of each reaction to the wells containing Hi-Di Formamide with LIZ size standards.


15.27.
15.28.
15.29.
15.30.
15.31.

Add only one sample per well.
Pipette up and down to mix.

Cap or cover the PCR plate.
Heat denature the samples at 95ºC for 3 minutes, then snap chill on ice for 5 minutes.
Prepare a sample sheet and injection list for the samples.
Run the samples on an ABI 3500 capillary electrophoresis instrument according to its user manual.
Review profile and controls, report results.

IGH + IGK B-Cell Clonality Assay

280314 Rev. H | October 2023
FOR RESEARCH USE ONLY; not for use in diagnostic procedures