I know the sequence of my gene but want to do the RACE to confirm the position of 5 end prime and 3' end prime. Now, I am confused about how RACE works in this case and we can say by seeing at results(RACE) that this is the start or end point of a gene?
Please help me to understand the RACE technique more precisely and its applications.
Thanks
Nicolas explanation above describes 3' RACE very nicely. However, it requires the presence of a 3' polyA tail. If you are sequencing a bacterial gene, this will not always be available. In this case you should ligate an adaptor primer to the 3' end and then proceed as described by Nicola. Regarding 5' RACE there are a number of interesting options which allows you to distinguish between true transcription start points and processed 5'ends. Here is a link to a paper in which they use this technique to identify the transcription start points of all transcripts in Helicobacter. In the paper there I seem to remember there is a link to their 5'RACE protocol
http://www.nature.com/nature/journal/v464/n7286/full/nature08756.html
good luck
Jesper
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