1 reply to this topic

[Epic]

Hello!!

Can someone define the use of -NE controls during PCR? What do you use for negatibe controls?
As negative controls I use 2:
RNA -ve (contains all cDNA components but not sample RNA)
RT-ve (contains all PCR components but not sample cDNA)

Cheers

[almost a doctor]

Epic, on Apr 14 2009, 05:00 PM, said:

Hello!!

Can someone define the use of -NE controls during PCR? What do you use for negatibe controls?
As negative controls I use 2:
RNA -ve (contains all cDNA components but not sample RNA)
RT-ve (contains all PCR components but not sample cDNA)

Cheers

Hello, I usually have 2 negative controls. They are slightly different from yours.

RT -ve control: this for me is actually all the RT reagents + RNA but NO RT enzyme. This way you have a control for presence of gDNA, and also show that on your RT +ve the reaction is happening.

NTC (no template control): this has all the PCR reagents but no DNA/cDNA (template). This will show that your PCR is specific and free of contamination.

If your RT -ve shows up in PCR: you have DNA in your RNA sample.

hope this helps