There one more kit for  5'/3'RACE  from Ambion that one also good

Hi Swati,

if you say Ambion do you mean the FirstChoice® RLM-RACE Kit? Just checking, with Ambion being part of Life Tech/Invitrogen being part of Thermo Fish.Sci., I don't find it that clearly distinguished anymore.

Thank you very much!

I used both Roche and Invitrogen RACE before, but there are as good as the Clontech SMART cDNA RACE kit. 

Hi,

I am using the Invitrogen kit for 5' RACE. I am getting smear with 3' nested primer and 5' Abridged anchor primer. I did the second round of PCR with nested primer and abridged universal amplification primer, but no luck. Please share your experience. Did you ( Anna-Lena Dittrich and Kai Wong ) able to get a discrete band?

Hi Swati, is your control reaction working or not?

Yes, my control reaction is working fine.

Are you expecting large transcripts?

You can try to use a touchdown PCR to amplify your second-round PCR. You may need to dilute your first-round PCR for the nested PCR.

But I think if you can perform the touchdown PCR for both rounds are even better.

Hello

i am also using the ROCHE 5/3 RACE kit, but i am having some issues.

The control reactions are not working, but i get a clear product with the expected size.

Also, i tried to sanger sequence my fragment and it succeed with the SP2 primer but not with the anchor primer. Is it possible to sequence with this primer?

Hi Sara,

I think, because the Anchor primer is made up of restriction endonuclease site sequence, so I don't think it is suitable to use for sequencing primer of the RACE PCR products. The best is to clone it into a vector for sequencing. Alternatively, we can design an inner primer to further sequence your PCR product.