MycoTOOL Mycoplasma Real-Time PCR Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
• The MycoTOOL Mycoplasma Real-Time PCR Kit is designed to test at least 10 cell culture samples for the absence of Mycoplasma.	• Automated workflow using automated sample preparation and Real-Time PCR • <5 hours, including sample preparation (data on file)

Upstream tips
• The MycoTOOL Mycoplasma Real-Time PCR Kit is designed to test at least 10 cell culture samples for the absence of Mycoplasma.
Protocol tips
• Automated workflow using automated sample preparation and Real-Time PCR • <5 hours, including sample preparation (data on file)

Publication protocol

A MycoTOOL mycoplasma real-time PCR kit was used in conjunction with a manual DNA extraction kit (QC sample preparation kit; Roche Diagnostics GmbH, Mannheim, Germany). All reagents necessary for gDNA extraction and real-time PCR are included in these kits (see supplementary Table 1). Sample processing, master mix preparation and setup for PCR amplification were performed in spatially segregated areas. Two separate PCR master mixes were prepared according to the kit instructions. The mycoplasma master mix contained primers and a fluorescein amidite-labeled detection probe for amplification and detection of mycoplasma gDNA and the PC plasmid. The RC master mix contained primers and a LightCycler yellow 555-labeled detection probe (Roche Diagnostics GmbH) for amplification and detection of the RC plasmid. MycoTOOL mycoplasma real-time PCR was performed in 96-well plates (Roche Diagnostics Corporation) at a volume of 50 μL/reaction according to the manufacturer's instructions. In each PCR plate, PC and test samples were run in technical replicates of four, with the no-template control in technical replicates of two. In each plate, five serial dilutions (GCs equivalent to 0 CFUs/mL, 0.1 CFU/mL, 1 CFU/mL, 10 CFUs/mL and 100 CFUs/mL) were used for M. arginini and four serial dilutions (GCs equivalent to 0 CFUs/mL, 1 CFU/mL, 10 CFUs/mL and 100 CFUs/mL) were used for M. hominis.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Cell Culture Contamination Detection Kit Mycoplasma using MycoTOOL Mycoplasma Real-Time PCR Kit from Roche Lifesciences.

Manufacturer protocol

Download the product protocol from Roche Lifesciences for MycoTOOL Mycoplasma Real-Time PCR Kit below.

Videos

Check out videos that might be relevant for performing Cell Culture Contamination Detection Kit Mycoplasma using MycoTOOL Mycoplasma Real-Time PCR Kit from Roche Lifesciences. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.