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
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Posted 14 April 2009 - 08:29 AM
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
