About 5' Race

[boyzoe]

Genome of my research species haven't sequenced yet. So after blast with other species, I designed primers and sequenced a fragment for the gene I researched in the species. Then I used 3' Race cloned the 3' fragment. However, I don't know how to get the sequence of 5'.
I know that Smart Race of Clontech can solve the problem. Is there other way to obtain 5' sequence of cDNA?
I propose no commercial kit will be used.
And if you guys have any experiences about 5', I hope you can talk of your method.
Thanks!

[JackBean]

What species are you working with?

What NA do you use? cDNA? I would suggest to do gDNA library, there you can perform 5' RACE, can't you?

[boyzoe]

What species are you working with?

What NA do you use? cDNA? I would suggest to do gDNA library, there you can perform 5' RACE, can't you?

The species is goldfish.

We haven't created gDNA yet. I cloned the gene from cDNA and sequenced 3' fragment. Blasted with zebrafish, there are almost 400bp to ATG. I want to obtain the full length of the gene.

Really grateful for your reply!

[JackBean]

you mean, that you miss 400bp of 5' part of gene?
Anyway, I think, even with cDNA this could work. Just design primer like in middle of gene and do the sequencing, you should get at least something
But otherwise, gDNA is the best choice in my opinion