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Why can't I just add Poly A to my Total RNA?
Hi all,
I have been weighing up my options with low input RNA. A bit of a debate has been going on regarding Poly A vs Total RNA in my lab. I would love to do Total RNA but I am not in love with the sample prep kits.
The options I am currently weighing up are Total RNA with TotalScript (I don't particularly love Nextera) and Poly A RNA with SMARTer (I particularly love this kit for amplifying the whole length of RNA and not using random primers).
Well, I had an idea that could let me have my cake and eat it too - what if I made all my RNA Poly A after Ribo Depletion and just used the SMARTer kit? It seemed so simple that I bet I am not the only person who thought of it which makes me think there is a reason why companies don't just do this.
What would be the drawbacks of converting my total RNA to Poly A and using a Poly A prep?
Help appreciated - thanks!
I have been weighing up my options with low input RNA. A bit of a debate has been going on regarding Poly A vs Total RNA in my lab. I would love to do Total RNA but I am not in love with the sample prep kits.
The options I am currently weighing up are Total RNA with TotalScript (I don't particularly love Nextera) and Poly A RNA with SMARTer (I particularly love this kit for amplifying the whole length of RNA and not using random primers).
Well, I had an idea that could let me have my cake and eat it too - what if I made all my RNA Poly A after Ribo Depletion and just used the SMARTer kit? It seemed so simple that I bet I am not the only person who thought of it which makes me think there is a reason why companies don't just do this.
What would be the drawbacks of converting my total RNA to Poly A and using a Poly A prep?
Help appreciated - thanks!