E value confidence

[magicsiew]

Hi, I have cloned out a gene that without annotation of my particular species in GenBank, when I blast the sequences, able to find the same gene from related species. But the e value are 0.074 and 0.006 respectively. By just analyzing from this data, can I conclude I have successfully clone out the gene? I think these e values are high, but since they are totally different species, so are these e value acceptable? Or is there any parameter of e value for different species?

[JackBean]

So, you have cloned your gene, sequenced and that sequence used in BLAST, rigth? From this you got 2 entries, with E-values 0.074 and 0.006.

So, what species are you talking about? At least, are they the same genus, order, kingdom(:lol)?
How long was your entry? What was the score and the coverage?
You didn't get any other hits?

[magicsiew]

I clone out 2 genes with 0.074 and 0.006 e value respectively. They are totally different species, banana against arabidopsis in Genbank database. My sequences are 850bp and 1kb in length.

The coverage are 2%, very small only, max score is 42.8 and max identical is 98%.

At 1st, I think of doing RACE, but since the coverage is too small, dont really have confidence on it. I try with FASTA and they show other product, rather than the 1 I found using BLAST.

[JackBean]

If there's only 2% coverage of 1 kb, that means only 20 similar nts!!! In such a case you cannot take that as significant (that short part may be, but not the whole region)

[magicsiew]

Yes. it cant... Thank you. Hmm... still trying with other methods...