Question: 5' RACE vs Random Hexamers or both
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gravatar for Mac
4.0 years ago by
Mac10
South Africa
Mac10 wrote:

I am trying to compare RACE and using Random Hexamers approaches..

(1) What is the best approach for creating cDNA from RNA, 5' RACE vs Random Hexamers or a protocol that uses both? (2) Is it possible not to use gene specific primers when making the cDNA (I want to use gene specific primers when applying the cDNA instead) and lastly, (3) How can I incorporate Primer ID; on cDNA synthesis or amplification if I am using the 5' RACE? 

next-gen • 898 views
ADD COMMENTlink modified 4.0 years ago by Devon Ryan92k • written 4.0 years ago by Mac10
1
gravatar for Devon Ryan
4.0 years ago by
Devon Ryan92k
Freiburg, Germany
Devon Ryan92k wrote:

These aren't really competing technologies, at least in my experience. One typically uses RACE to find the UTR of specific transcripts. One uses random hexamers to amplify everything. So the question isn't so much "which approach is better", but rather "which approach is appropriate for whatever I want to do". You can certainly just make cDNA of one transcript (or gene, depending on how it's spliced), it's just a matter of whether you want to go through the hassle of designing and order the correct primer instead of just using an off the shelf cDNA kit.

ADD COMMENTlink written 4.0 years ago by Devon Ryan92k

Thanks Devon!

ADD REPLYlink written 3.9 years ago by Mac10
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