RNAi showing upregulation via QPCR
Posted 09 March 2011 - 01:33 AM
But when we come to analysing mRNA levels via QPCR we consitently get a fold increase in the range of 600-1000 times more in the cells treated with dsRNA.
Initially we thought this could be due to the QPCR primers amplifying our dsRNA this is not the case as they are both targeted to seperate regions of the gene. We know that the knockdown is having the correct effect via phenotypic analysis but this is a bit puzzling.
Has anyone seen this before or can anyone provide some insight as to what may be happening and what can be done to correct this?
Posted 10 March 2011 - 01:45 PM
A 1000 fold increase can only come from spiked external something.
Posted 12 March 2011 - 11:26 PM
Posted 24 August 2011 - 03:02 PM