A question for Reverse transcription experts
Posted 25 April 2013 - 02:13 PM
I Have RNA isolated from murine macrophage. I want to creat cDNA. I have a kits from invitrogen that include no primers and no obvoius polyA tail polymerase Enzyme.
. The question is
Can the reverse transcriptase E act as ploy A polymerase at the same time under the same temperature.
Can I use the miRNA primers to creat cDNA for the miRNAs only but not all RNA ? is it critical to have an random primers or what
If I do this cDNA synthesis and I want to be sure that my product (total RNA ) get changed to cDNA, what is the accurate method for confirming this ? Can I measure the cDNA concentration after RT or this is not indicative ?
Posted 25 April 2013 - 04:11 PM
Measuring the DNA content after transcription doesn't work as the dNTPs and any residual RNA will also give a reading on a spec.
Posted 25 April 2013 - 09:02 PM
Posted 26 April 2013 - 05:10 AM
Posted 26 April 2013 - 07:34 AM
Posted 26 April 2013 - 01:57 PM