eamag/protocols-with-steps huggingface dataset:
TITLE: MicroRNA Affymetrix microarray probe labeling from ocular fluids
AUTHORS: Zeljka Smit-Mcbride

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Sample collection, isolation of miRNAs, and quality control </span></div><div class = "text-block"><span>Aqueous and vitreous humor, as well as plasma samples, have been collected from Diabetes mellitus (DM) and control patients during the standard-of-the-care eye surgery. Samples of 100-200ul were collected, aliquoted in 100ul aliquots, frozen on dry ice immediately and stored at -80</span><span style = "vertical-align:super;">o</span><span>C. RNA was isolated using Exiqons’ modification of Qiagen’s microRNeasy kit. Quantification and quality check of isolated miRNAs was performed on BioAnalyzer (Agilent) with Small RNA microfluidics Chips. </span></div><div class = "text-block"><span style = "font-weight:bold;">Affymetrix microarray probe labeling</span></div><div class = "text-block">Probe labeling was done from 10 ng of aqueous, 20ng of vitreous and 20ng of plasma miRNA samples from a total of 24 aqueous, 24 vitreous and 25 plasma samples using FlashTag (Affymetrix) procedure. Labeled probes were hybridized to Affymetrix miRNA Array 3.0 in UC Davis Genome Center Microarray Core Facility using the standard procedure (Affymetrix, Santa Clara, CA). The total data set included 73 Affymetrix miRNA 3.0 microarrays. Upon scanning of the hybridized and washed Chips, data was obtained in a set of Affymetrix data files (.cel, .arr, .jpg). </div></div>

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This is the protocol for the PhireKit touchdown PCR assay</p>

[BEFORE_START]
<p>The assay has been optimized and validated for the Bio-rad C1000 Touch Thermocycler.</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
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?.
TITLE: S. O. C. medium
AUTHORS: Andreas Sagen

[DESCRIPTION]
S.O.C. Medium is used in the final step of bacterial cell transformation to obtain maximal transformation efficiency ofE. coli. S.O.C.

[STEPS]
SECTION: 500 mL 1 M Glucose solution
1. Fill the bottle with 400 mL distilled water
SECTION: 500 mL 1 M Glucose solution
2. Measure 90.08 g Glucose

Materials:
 
SECTION: 500 mL 1 M Glucose solution
3. Add distilled water to a total of 500 mL
SECTION: 500 mL 1 M Glucose solution
4. Filter sterilize with a 0.2 µm filter
SECTION: 1 000 mL S. O. B. medium
13. Fill the bottle with 950 mL distilled water
SECTION: 1 000 mL S. O. B. medium
14. Measure 20 g Tryptone, 5 g Yeast extract and 0.5 g Sodium chloride. Add 15 g Agar to create agar plates

Materials:
 
 
 
 
SECTION: 1 000 mL S. O. B. medium
16. Adjust pH to pH 7.0 with concentrated sodium hydroxide

Materials:
 
SECTION: 1 000 mL S. O. B. medium
17. Add distilled water to a total of 1000 mL
SECTION: 1 000 mL S. O. B. medium
18. Autoclave solution at 121 °C for  
SECTION: 125 mL 2 M Magnesium chloride
9. Fill the bottle with 100 mL distilled water
SECTION: 250 mL 250 mM Potassium chloride
5. Fill the bottle with 200 mL distilled water
SECTION: 100 mL S. O. C. medium
19. Sterilely transfer 98 mL S. O. B. medium to a sterile 100 mL container
SECTION: 100 mL S. O. C. medium
20. Sterilely add 2 mL 1 Molarity (M) Glucose solution
SECTION: 250 mL 250 mM Potassium chloride
7. Add distilled water to a total of 250 mL
SECTION: 250 mL 250 mM Potassium chloride
6. Measure 4.66 g Potassium chloride

Materials:
 
SECTION: 250 mL 250 mM Potassium chloride
8. Autoclave solution at 121 °C for 15 min

SECTION: 125 mL 2 M Magnesium chloride
12. Autoclave solution at 121 °C for 15 min

SECTION: 125 mL 2 M Magnesium chloride
11. Add distilled water to a total of 125 mL
SECTION: 125 mL 2 M Magnesium chloride
10. Measure 50.825 g Potassium chloride

Materials:
 
SECTION: 1 000 mL S. O. B. medium
15. Add 1 mL Potassium chloride solution (250 millimolar (mM)) and 0.5 mL Magnesium chloride solution (2 Molarity (M))
TITLE: Doctor Vida Pocket - COVID-19 assay - Instructions for Use
AUTHORS: goncalo.doria 

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Instruction of use for Doctor Vida pocket - COVID-19 assay</div></div>

[STEPS]
?. [Preparation setup]
Turn on Dr. Vida Pocket device - by plugging the Dr. Vida Pocket device's power supply into a suitable power plug (120V/240V AC) and connecting the micro-usb plug into the Dr. Vida Pocket device.
When turned on, the device will emit a short bip.
?. [Start new assay]
Press "Start new assay" in the Dr. Vida pocket mobile App (under Assays menu)
?. [Preparation setup]
Login into Dr. Vida pocket mobile App (Android / iOS) and connect the app to the device (under Devices menu)
?. [Start new assay]
Scan the Data matrix of Dr. Vida pocket - COVID 19 assay reaction tube (or, alternatively, fill in the assay Reference and Lot ID, as supplied in the box), and press Proceed
?. [Start new assay]
Insert Sample ID and press Proceed
?. [Results]
After  minutes, the device will issue a long bip indicating that the assay has finished and that the data is ready for transfering. Make sure that the Dr. Vida pocket app is still running and the device is connected to allow the data transfer.After data transfer, the device issues a short bip and the final result will appear in the app screen.
In case you wish to access more details about this and past assay results, please consult assay results under Results menu.
Detected: This indicates the presence of the SARS-CoV-2's RNA in the collected sample Not detected:  This indicates that the assay did not detected SARS-CoV-2's RNA in the collected sample, considering its estimated LoD of ^.Failed: This indicates that the assay was not sucessfull. Please repeat the assay.
?. [Assay processing]
Click Proceed in the Dr. Vida pocket app and follow the instructions to add the Reaction tube inside the device.
Do not open the cap of the device during the assay. The temperature inside the device reaches
?. [Sample collection]
Collect a swab (nasopharyngeal and/or oropharyngeal) from the individual to be tested following the local collection guidelines.
?. [Sample collection]
Insert swab inside the provided Collection tube (containing  ) and spin the swab for  seconds.
[collection buffer]
?. [Sample collection]
Remove the swab and discard it properly into a biohazard disposable container, following local biohazard waste procedures.
?. [Sample collection]
Insert the provided minivette inside the Collection tube and wait a few seconds until the sample liquid reaches the minivette's white filter. If necessary, tilt the tube 45º to help the capillarity filling.
During this step, make sure not to block the hole on top of the white part of the minivette.
?. [Sample collection]
Remove the minivette from Collection tube and dispose its content ()  inside the Reaction tube.
10 µl
Make sure that the tip of the minivette is properly inserted until the bottom of the Reaction tube and that you block the hole on top of the white part of the minivette while disposing its content.
TITLE: Amplifying the target genomic region by PCR
AUTHORS: Hanqin  Li, Yogendra Verma, Dirk Hockemeyer, Frank Soldner

[DESCRIPTION]
This protocol describes a procedure for amplifying targeted genomic regions using a PCR reaction


[STEPS]
1. For a 150bp paired-end or single-read sequencing experiment, design primers in a way that the region of interest, like CRISPR cutting site or specific mutations, locates within 130bp from at least one of the primers
2. Add NGS adapters to 5’ end of primers following the instructions of your local NGS facility
3. 20 µl PCR reaction for each sample
3.1.  PCR Reaction - setup
     Ultrapure H2O      10.6µl        5x HF buffer      4µl        2.5mM dNTP       1.6µl        10uM primer Forward      0.5µl        10uM primer Reverse      0.5µl        DMSO      0.6µl        Titan DNA polymerase   or Phusion      0.2µl        Crude cell lysis      2µl   
 
4. Calculate the amount of each component needed for all samples, also count a negative control and positive control
5. In a pre-PCR area, mix all components except the crude cell lysate to a microcentrifuge tube or a 15 ml conical tube to make master mix
6. Aliquot 18 µl to each 200 µl microcentrifuge tube or each well in a 96-well PCR plate. Use reservoir and multi-channel pipet if there are many samples.
7. Add 2 µl crude cell lysate to the reaction. In one reaction, use 2 µl H2O instead as negative control. In another reaction, use 2 µl previously validated crude cell lysate as positive control.
8. Cap the tubes or seal the plates properly with an adhesive seal.
9. Shake the tubes or plates vigorously to mix
10. Briefly spin the samples 
11. Use the following touch-down PCR protocol in a thermocycler
11.1.  Touch-down PCR protocol
     98C      3min        98C      30s        70C (touch down,   1C/cycle)      30s        72C      30s        Go to 2       12 cycles in total        98C      30s        58C      30s        72C      30s        Go to 6       23 cycles in total        72C      7min        4C or 12C      forever
TITLE: Two-Bottle Choice Protocol
AUTHORS: Marcella Hughes, Olivier George

[STEPS]
?. [24-Hour Two Bottle Choice Procedure]
Record weekly body weights of animals for calculation of solution intake as mg/kg
?. [24-Hour Two Bottle Choice Procedure]
Mix and fill required solutions in bottles
?. [24-Hour Two Bottle Choice Procedure]
Record weight of all bottles prior to drinking session
?. [24-Hour Two Bottle Choice Procedure]
Put bottles on cages with solution A on left and B on right (rotate side position each day) and leave for 24 hours
?. [24-Hour Two Bottle Choice Procedure]
Following 24 hours remove bottles from caging being careful not to spill any liquid
?. [24-Hour Two Bottle Choice Procedure]
Record weight of all bottles after drinking session
?. [24-Hour Two Bottle Choice Procedure]
Subtract weight of each bottle after drinking from weight of same bottle prior to drinking to calculate liquid consumed and record in notebook
?. [24-Hour Two Bottle Choice Procedure]
Rinse bottles and re-mix and fill solutions in bottles
?. [24-Hour Two Bottle Choice Procedure]
Record new weights and place bottles back on cages with positions swapped
?. [24-Hour Two Bottle Choice Procedure]
Repeat all steps daily as desired
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>We recently developed a continuous flow system to transform microorganisms in high throughput in a microfluidic device (Garcia et al., 2017). This system employs microfluidic channels that contain a bilateral constriction between the inlet and outlet electrode connections (<em>length</em> = 3.0 mm, <em>width</em><sub>min</sub> = 50 μm, <em>width</em><sub>max</sub> = 2.0 mm, and <em>height</em> = 100 μm). The constriction amplifies the electric field under an applied voltage between the inlet and outlet electrodes to levels sufficiently high to induce electroporation. During <em>P. caudutus</em> transformations, the cells were driven through the microfluidic device at flow rates of 50 μL/min and 500 μL/min, which correspond to residence times (i.e. pulse durations) of 20 ms and 2 ms, respectively. Square wave pulses with, for example, 5 ms ON and 5 ms OFF cycles (50 % duty cycle) are applied to the microchannel through the dispensing needle. Therefore the cell viability cannot be accurately evaluated since only 50 % of the cells experience the electric field. The pulses are delivered from electrodes with alternating polarity between the pulses to reduce electrolytic effects at the electrode-buffer interface (Fig. 1). After flowing through the microchannel, each 200 μL cell sample is added to a 1.5 ml Eppendorf tube containing 1 ml of fresh growth media for cell recovery. The applied voltages we evaluated had amplitudes of 250 V (<em>E<sub>max</sub></em> = 1,500 V/cm), 375 V (<em>E<sub>max</sub></em> = 2,250 V/cm), and 500 V (<em>E<sub>max</sub></em> = 3,000 V/cm) for each polarity. The non-uniform constriction in the microfluidic devices generates a variable electric field that is capable of transfecting cells while minimizing exposure to the highest electric field. </p>
<p> </p>
<p><strong>Figure 1</strong>: Electric field waveforms employed for transient and stable transfection of <em>Bodo caudatus</em>. Three independent electroporation systems were used for reproducible transfection including a) our microfluidic electroporation platform (Garcia et al., 2017), the NEPA21 square-wave transfection system (BulldogBio), and the MicroPulser<sup>TM</sup> exponential decay electroporator (Bio-Rad). Additionally, the b) signature waveforms for the NEPA21 square wave transfection system of ‘poring’ and ‘transfer’ pulses for electroporation are shown. Note: The time scale in Fig. 1a is a zoomed-in version of the red-dashed box from Fig. 1b.</p>
<p> </p>
<p>Garcia, P.A., Ge, Z., Kelley, L.E., Holcomb, S.J., and Buie, C.R. (2017) High efficiency hydrodynamic bacterial electrotransformation. <em>Lab Chip</em> (<strong>17</strong>): 490-500.</p>

[STEPS]
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TITLE: jres_metab.nan
AUTHORS: NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison

[DESCRIPTION]
This is a protocol for running the Bruker pulse program "jresgpprqf".

[BEFORE_START]
This protocol assumes your sample is loaded, locked, tuned, and shimmed in the magnet.

[GUIDELINES]
This protocol intends to provide concise instructions to carry out the experiment. For more comprehensive information, see Bruker's documentation "Basic NMR Experiments" by clicking ? →  Manuals (docs) on the menu bar on TopSpin. See also "Pulse Program Catalogue. 1D/2D" for the details about the pulse program used in this protocol.

[STEPS]
SECTION: Create a new dataset
1. 
SECTION: Create noesypr1d experiment file
0.22. When Lock, Tune, Shim are complete proceed to the specific protocol for your desired pulseprogram(s).
SECTION: Create a new dataset
1.1. On the menu bar on TopSpin, click on
Start → Create Dataset
 
 

SECTION: Create a new dataset
1.2. Enter
NAME: Name of a set of datasets (e.g., human_serum_study1). Use a single string (do not use spaces)
EXPNO: Dataset number. Use a positive integer
 
Select
Directory: Your directory 

 

SECTION: Create a new dataset
1.3. Select
Read parameterset

Click the button
Select
SECTION: Create a new dataset
1.4. A new window opens. On the right top bar, select
Source = /opt/NAN_METAB/par 
 
In the list, select the one you want to use:
JRES_br600_serum.par: Parameter set optimized for serum and plasma samples.
JRES_br600_urine.par: Parameter set optimized for urine samples.
 

Click
OK
SECTION: Create a new dataset
1.5. Click
OK
SECTION: Acquire a spectrum
2. Go to the "USE DEFAULT" tab below to proceed with the default optimized parameters.
SECTION: Acquire a spectrum
3. 
SECTION: Acquire a spectrum
3. 
SECTION: Acquire a spectrum
3.2. Click on
Acquire → Gain
in the menu bar to automatically set the receiver gain.
 
 

SECTION: Acquire a spectrum
3.3. Click
Go
in the menu bar to acquire a spectrum.

 

SECTION: Acquire a spectrum
3.4. After the run, click on
Process → Proc. Spectrum
in the menu bar to execute an automated processing macro.
 

SECTION: Acquire a spectrum
3.5. If you want to modify parameters to improve your spectrum,   and move to the step case "MODIFY PAR".

SECTION: Acquire a spectrum
4. Acquire a spectrum. If more optimization is needed,  .

 

SECTION: Acquire a spectrum
3.1. Select the AcquPars tab to show a parameter set.
 
 

SECTION: Acquire a spectrum
3.2. Click the icon
_∏_
to see a complete parameter set.

 

SECTION: Acquire a spectrum
3.3. Change parameters as needed.

Typically, the main parameter to adjust is NS:
 

SECTION: Acquire a spectrum
3.4. Change parameters as needed.

Common parameters to adjust:
 

 

SECTION: Acquire a spectrum
3.1. Load the calibrated P1 using the following command in the command line.
 
(e.g., getprosol 1H 10.01 -7.45)
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol describes how to perform PCR on extracted <em>Symbiodinium</em> DNA to amplify a fragment of the chloroplast 23S rRNA gene. It is based on the method describes in Pochon <em>et al.</em>, 2006. The protocol then explains how to perform an RFLP (restriction fragment length polymorphism) analysis on the PCR products as well as eitherusing them directly for sequence analysis by Sanger sequencing or cloning them into a plasmid back bone to separate genes from mixed <em>Symbiodinium</em> populations and perform the sequence analysis on several clones per original sample.</p>
<p> </p>
<p>The attached file contains reference sequences of the following Symbiodinium strains: Clade A strains SSA01, SSA02, SSA03; SSB01 (clade B), SSB01del6, SSB01del136, SSE01 (clade E) and A001 (Clade D).</p>

[BEFORE_START]
<p><strong>Have/order the primers for the <em>Symbiodinium</em> cp23S rDNA locus:</strong></p>
<p> </p>
<p><strong>Primer sequences:</strong></p>
<p><strong>23S4F (forward): 5’-GAC GGC TGT AAC TAT AAC GG-3’</strong></p>
<p><strong>23S7R (reverse): 5’-CCA TCG TAT TGA ACC CAG C-3’</strong></p>

[GUIDELINES]
<p><strong>20x Borax running buffer for agarose gels:</strong></p>
<ul>
<li>Dissolve 38.18 g Sodium Tetraborate Decahydrate (Borax) in 900 ml distilled water (you might need to heat up the solution to dissolve everything)</li>
<li>Bring to 1 L with distilled water</li>
<li>DO NOT try adjusting the pH, it takes a looooot of boric acid, even with directly adding pure powder and the gels are running well without doing it.</li>
</ul>
<p>This results in a 100 mM solution, adjust the amount accordingly, when using other forms of Tetraborate (anhydrous etc.).</p>
<p> </p>
<p>Add 50 ml of the 20× buffer to 950 ml distilled water to obtain a 1× solution.</p>
<p> </p>
<div><a href="https://www.future-science.com/doi/abs/10.2144/04362BM02" target="_blank" rel="noopener noreferrer">Sodium boric acid: a Tris-free, cooler conductive medium for DNA electrophoresis</a></div>
<div>Jonathan R. Brody and Scott E. Kern</div>
<p>BioTechniques 2004 36:2, 214-216</p>

[STEPS]
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TITLE: CRISPR puromycin 
AUTHORS: Rebecca Berrens

[DESCRIPTION]
This protocol is for CRISPR knock-in generation in mouse embryonic stem cells

[BEFORE_START]
Have ES cells ready growing happily. Mostly 2 passages after thawing

[STEPS]
1. Plate cells on 6-well plate on SNL a day before transfection (number of cells shown in the table below).
3. A day after transfection cells were passaged from 6-well plate into 10cm Petri dishes with SNL. Cells from each well were split into 3 Petri dishes in 3 different densities (1/2, 1/3 and 1/5 of the cells).
4. 24 hours later apply puromycin selection (at the adequate concentration for your cell line of interest) for 48hours to select for cells that contain the Cas9 plasmid (there is no selection in the targeting vector!).
5. After 48 hours of puromycin selection feed cells with regular ES medium every day until ready to pick colonies (usually 12-14 days after transfection).
6. pick colonies (usually 12-14 days after transfection).
follow ES cell picking protocol
2. Transfect cells using Lipofectamine 2000 protocol and plasmid DNA amounts as shown in the following table: 
 
E14 1x10^6 cellsE14 1x10^6 cells6:1 Molar Ratio dTAG-SETDB1 to MA299/300No DNA Control100uL OPTIMEM + 6.46 uL (2.5 ug) dTAG-SETDB1 + 0.44 uL (869.3ng) MA299/300100ul OPTIMEM100uL OPTIMEM + 10 uL LF2000100uL OPTIMEM + 10uL LF2000
TITLE: ssDNA2.0: Ligation mix II
AUTHORS: Sarah Nagel, Anna Schmidt, Matthias Meyer

[DESCRIPTION]
Protocol for the preparation of Ligation mix II for automated single-stranded DNA library preparation using the ssDNA2.0 method (Gansauge et al. 2020). 

References
Gansauge, M.-T., Aximu-Petri, A., Nagel, S., & Meyer, M. (2020). Manual and automated preparation of single-stranded DNA libraries for the sequencing of DNA from ancient biological remains and other sources of highly degraded DNA. Nature Protocols, 15, 2279-2300.

[STEPS]
TITLE: Working Theory of Flow Cytometry
AUTHORS: 181830691 

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Flow Cytometry (FCM) is a scientific technology developed in the 1970s. In the 1980s, its application rangewas developing from basic research to clinical medical research and disease diagnosis and treatment monitoring. It set of optics, electronics, fluid mechanics, cell chemistry, biology, immunology. At the same time, it has the function of analyzing and sorting cells, and can quickly analyze the cell suspension. The light scattering and fluorescence indices of the cells were obtained by detecting the single row of cells in the flowing liquid one by one.</div></div>

[STEPS]
TITLE: Subcellular Fractionation
AUTHORS: Szu-Chi Liao, Mohamed Taha Moutaoufik, Ken Nakamura

[DESCRIPTION]
To isolate total cell lysate into cytosolic and mitochondrial, soluble vs insoluble fractions.

[STEPS]
1. Harvest cells with a scraper.
2. Resuspend PBS and centrifuge at 500g for 5 min.
3. Homogenize using a glass pestle in isolation medium.
4. Centrifuge at 600 g for 10 min at 4°C.
5. Transfer the supernatant to a new microcentrifuge tube and centrifuge at 9,000 g for 10 min at 4°C.
6. Transfer the supernatant (cytosolic fraction) to a new microcentrifuge tube.
7. Resuspend the pellet with isolation medium.
8. Centrifuge at 9,000 g for 10 min at 4°C.
9. Repeat step 7 and 8 for a total of 3 times.
10. The final pellet is collected as the mitochondrial fraction.
11. Add Triton X-100 to both cytosolic and mitochondrial fractions to a final concentration of 1%.
12. Incubate mixture from both fractions on ice for 30 min.
13. Centrifuge at 20,000 g for 10 min at 4°C.
14. Transfer the supernatant (soluble fraction) to a new microcentrifuge tube.
15. Incubate the pellet in SDS buffer for 30 min.
16. Centrifuge at 20,000g for 10 min at room temperature.
17. The final solution is the insoluble fraction.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Protocol for seamless, scarless, homology-based cloning of vectors using an enzyme-free technique. </p>
<p>The method was published by Zurbriggen et al., 2015, doi: <a href="https://dx.doi.org/10.1371%2Fjournal.pone.0137652" target="_blank">10.1371/journal.pone.0137652</a></p>
<p>It relies on the endogenous homologous recombination machinery of <em>E. coli</em> lab strains.</p>

[BEFORE_START]
<p>Make sure to recover high concentrations of fragments.</p>

[STEPS]
?. 
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TITLE: Zebrafish (Danio rerio) Environmental Summary, Aquatic Resources Program, Boston Children&#39;s Hospital 2020a
AUTHORS: Christian Lawrence, Jason Best, Althea James, Shane Hurley, Mitchel Shia, Michelle Urh, Brady Hirshfeld, Nina Bakker, Hugo Perdomo, Aaron Krueger

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Environmental and housing conditions experienced by laboratory animals exert profound effects on their biology, physiology, and behavior.  These parameters are important and often overlooked sources of potential variation in experiments, and should be reported in peer-reviewed publications in order to promote scientific reproducibility.  To that end, here we provide a summary of the environmental conditions in zebrafish (Danio rerio) aquaculture facilities at Boston Children's Hospital (BCH).   We include data on the physico-chemical, health, and nutrition of zebrafish in three separate facilities at BCH: Karp, Enders SSB, and Enders Lobby. In this year's version, we also include new information on our feeding practices and colony pathogen history. This information will be applicable to any study involving zebrafish conducted in one of these three facilities during the year of 2020.</div></div>

[STEPS]
?.
TITLE: NHS-ester-protein-labeling
AUTHORS: Liv Jensen

[DESCRIPTION]
Protocol for labeling a purified protein with an NHS ester fluorescent dye.

[STEPS]
1. Mix 40μM unlabeled protein with 80μM ATTO 565 NHS ester dye in labeling buffer.
2. Incubate 1 hr at room temperature.
3. Buffer exchange the reaction into quench buffer over a pre-equilibrated G-25 desalting
column.
4. Assess labeling efficiency by measuring the ratio of absorbance at 280 and 564 nm,
correcting for dye absorbance at 280nm, using a Nanodrop spectrophotometer