i'm carrying out gene expression experiment on bacteria. i used 16sRNA as a refrence gene, but the Ct value is around 6 while one of my target gene Ct is 27, should i start with a lower cDNA conc. and shift the target gene Ct >30, decrease the primer concentration for the refrence gene or accept the result as it is. does anyone have a clue.
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#2
Trof
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Posted 12 May 2011 - 05:09 AM
Decrease primer concentration for reference gene. It's called primer limiting. The Ct of your gene of interest and reference gene should be similar. You may try less abundant reference genes.
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#3
lilies
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Posted 13 May 2011 - 09:13 AM
Trof, on 12 May 2011 - 05:09 AM, said:
Decrease primer concentration for reference gene. It's called primer limiting. The Ct of your gene of interest and reference gene should be similar. You may try less abundant reference genes.
