help - (Aug/18/2007 )
QUOTE (methylnick @ Nov 22 2007, 01:24 PM)
yongyi, if you are using the conventional method, you won't be able to see a band after conversion, because you would have much less converted DNA than what you started with. you need to perform the PCR and look for the amplicon.
Nick
Nick
Nick:
Thank you very much! But the problem is that I can't see the band after PCR.I think it is the problem of the my primer . So I want to design the primer again.I want to study the promoter of hoxc8 gene of sheep but i can't find it .I find the promoter of human's hoxc8,so i want to design the primer use it. But i can't design the appropriate primer,the primer that i designed didn't work.Could you help me design ?Thanks in advance!
-yongqi_1-
yongyi,
without knowning the sequence of hoxc8 in sheep it maybe difficult, I am not too sure how diverged this sequence is compared to chicken, or even cow?
the first thing to do would be to sequence hoxc8 in sheep before looking at methylation by bisulfite, I think the primers you havepicked may not work in sheep if they were based on chicken sequence.
once you know the genomic sequence of sheep hoxc8 then you can go ahead and design primers for bisulfite to it.
it's a little difficult to work on a species where the genome is not currently complete!! 
-methylnick-
QUOTE (methylnick @ Nov 30 2007, 12:12 AM)
yongyi,
without knowning the sequence of hoxc8 in sheep it maybe difficult, I am not too sure how diverged this sequence is compared to chicken, or even cow?
the first thing to do would be to sequence hoxc8 in sheep before looking at methylation by bisulfite, I think the primers you havepicked may not work in sheep if they were based on chicken sequence.
once you know the genomic sequence of sheep hoxc8 then you can go ahead and design primers for bisulfite to it.
it's a little difficult to work on a species where the genome is not currently complete!!
without knowning the sequence of hoxc8 in sheep it maybe difficult, I am not too sure how diverged this sequence is compared to chicken, or even cow?
the first thing to do would be to sequence hoxc8 in sheep before looking at methylation by bisulfite, I think the primers you havepicked may not work in sheep if they were based on chicken sequence.
once you know the genomic sequence of sheep hoxc8 then you can go ahead and design primers for bisulfite to it.
it's a little difficult to work on a species where the genome is not currently complete!!
nick:
Thank you very much! Thanks for your help!
-yongqi_1-