USER MANUAL

Cat. No. 093050401 (25 tests)

Myco-Sniff-Rapid

Cat. No. 093050402 (50 tests)

TM

Revision Date: 2021-08

Mycoplasma Luciferase Detection Kit

STORAGE

Collect Cell Culture-20 °C in the dark for 1 year. Reconstituted Myco-Sniff-Rapid™ Reagent andSubstrate can be stored at -80 °C for six months, at -20 °C for one month, orat 2-8 °C for one week. Repeated freezing and thawing up to 10 times will notaffect the accuracy of the test results.

Culture the cells for at least 24 hours, collect the cell culture medium andcentrifuge at 400 × g for 3 minutes. The supernatant should be tested
immediately or stored at 2-8 °C for no more than one week. Avoid freezing
and thawing the collected medium.
NOTE For optimal assay performance, cell confluency should reach 80% or higher.

DESCRIPTION
Myco-Sniff-Rapid™ Mycoplasma Luciferase Detection Kit offers a quick (20 mins),simple and sensitive method for detecting mycoplasma contamination in cellcultures. This innovative kit utilizes the activity of mycoplasma metabolic
enzymes from 209 mycoplasma species, including those commonly foundcontaminating cell cultures. Since this enzyme is not present in eukaryoticcells, the test is specific for mycoplasma. In the presence of mycoplasma, theenzymes react with the substrate to catalyze the conversion of ADP to ATP.
Mycoplasma contamination can be detected by measuring the level of ATP ina sample before and after the addition of Myco-Sniff-Rapid™ substrate via aluciferase assay.

BENEFITS� Rapid results in 20 minutes
� Highly sensitive� Specific for detecting all common mycoplasma contaminant species
� No interference with cell culture media components
� Simple operation - add reagents directly to the culture supernatant
093050401(25 tests)

093050402(50 tests)

Myco-Sniff-Rapid Reagent (lyophilized)

2 vials

4 vials

Myco-Sniff-Rapid Substrate (lyophilized)

2 vials

4 vials2 x 1.5 mL

4 x 1.5 mL

Components

Myco-Free Water

Assay (avoid bright lighting)1

Equilibrate the dissolved Myco-Sniff-Rapid Reagent, Myco-Sniff-Rapid
Substrate and cell culture medium supernatant to room temperature.

2Add 50 μL Myco-Sniff-Rapid Reagent and 50 μL cell culture medium
supernatant to a 1.5 mL tube or 96-well plate and incubate at room
temperature for 5-10 minutes.
NOTE Mix the samples gently with a pipette and be careful not togenerate any large bubbles. Small bubbles on the edge of the tube or wellshould not impact assay result.

3

Place the tube or 96-well plate in a luminometer to measure theluminescent signal value (Reading A).

4

Add 50 μL of Myco-Sniff-Rapid Substrate to the tube or 96-well plateand incubate at room temperature for 10-15 minutes.
NOTE Mix the samples gently with a pipette and be careful not togenerate any large bubbles. Small bubbles on the edge of the tube orwell should not impact assay results.

5Place the tube or 96-well plate in a luminometer to measure theluminescent signal value (Reading B).

6

Data interpretation: The ratio of Reading B to Reading A is used todetermine whether a cell culture is contaminated by mycoplasma.

PROCEDURE

Ratio(Reading B/Reading A)
B/A ≥ 1

Mycoplasma contamination

B/A ≤ 0.8

Negative for mycoplasma

Materials Required, But Not Supplied
96-Well plate, Luminometer (tube luminometer, plate luminometer or
multifunctional reader), Pipette, Pipette tips, Centrifuge

Component Reconstitution

Results

0.8 < B/A < 1Samples should be retested after another24-48 hours culture in quarantine. If the B/Aratio remains between 0.8 and 1 with nosignificant increase, the sample is negative
for mycoplasma.

Reconstitute one vial of lyophilized Myco-Sniff-Rapid Reagent and one vialof Myco-Sniff-Rapid Substrate by adding 700 μL Myco-Free water to each.
Wait until rehydration is completed.

MP BIOMEDICALSAMERICAS: 800.854.0530 | custserv.na@mpbio.com
EUROPE: 00800.7777.9999 | custserv.eur@mpbio.com
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