RACE kits - opinions (View forum version)


Posted 19 February 2010 - 05:16 AM

I am trying to get the full sequences of several RNA within Drosophila, having only a very small amount of starting sequence in the middle of each gene (I've verified expression with RT-PCR so the RNA should be there). I would therefore like to do 3' and 5' race on multiple genes. I was thinking of using Invitrogen's RLM RACE kit (generacer) or BDclontech's SMART RACE kit to prepare my cDNA.

It seems the technologies they use are slightly different, but both lead to the same end product (full length cDNA upon which 3' and/or 5' RACE can be performed). It looks like generacer has a much longer protocol for preparing the cDNA because you have to use CIP and TAP to modify the 5' end, whereas SMART RACE's technology doesn't require these steps - does the extra work in the invitrogen kit produce better/more specific results? I've also seen the ambion kit which has generally the same protocol as invitrogen.

I am wondering how much success any of you have had with any of these kits (ideally if someone has used multiple that would be the best). If there's another kit you have used with success please let me know that also! Since I'm working with many genes and want to do 5' and 3' RACE on each I think I will be sticking with a kit that produces RACE-ready cDNA, rather than one that uses a GSP in the initial RT step for 5' RACE.

Thanks in advance


Posted 22 February 2010 - 08:16 AM



Posted 23 February 2010 - 02:57 AM

we´ve used ClonTech - no probs.
don´t know about invitrogen.