Gene Expression Issues
Posted 08 August 2011 - 07:43 AM
Posted 10 August 2011 - 12:04 PM
Posted 10 August 2011 - 02:32 PM
I don't understand your problem. Your question would suggest that 18s is the highest expressing gene in a cell? You certainly could have a target sequence with a higher cT value than an 18s target (assuming you load the same amount of mRNA for both)
But doesn't rRNA make up >80% of the RNA in cells? Since 18s is a rRNA, I thought that it would have a very high expression. Could the Alu sequences have higher expression than the rRNA? Does the 293t cell line contain less rRNA than other cells?
Posted 11 August 2011 - 01:25 AM
Other thing may be, if your Cts are really low (< 15), that 18S is so abundant it inhibits the reaction. If you dilute your sample like 500 - 1000x you may get Ct values in the safer range.
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