Those multiple bands may be due to the presence of more than 1 restriction sites for that enzyme. it may also due to star activity of that enzyme. so it will be useful to check the sequence and the enzyme activity.

Hi Neelofar

if you can load the photograph of autoradiogram...it would be nice...if you have some background information about the gene organization and restriction map of the gene of your interest...it could be easy to interpret your results.....

Guruprasad has mentioned some improtant points above.....also keep in mind the partial digestions and sensitivity of restriction enzymes to methylation etc...

additionally ....If possible you can also make use of q-PCR to find out the copy number of your gene in the genome using either a standard cure approach or using a known reference gene which has a single copy in the genome....

bye

Hi! You are telling to find out copy number of gene.

First what is your gene, how many restriction cites it has? You have digested at a time with many enzymes obviously you see different number of bands. Out of those enzymes how many are rare cutters and how many are frequent cutters. Then it is very difficult to say. Because of sequence duplication sometimes you see many bands even if you digest with single RE.

Hybridization temp. also play important role in specificity of hybridization.

So dear first sit coolly and plan you work, go as per that. Refer literature now days you get everything through web searching, provided if you have internet facility. Wikipedia also helps a lot.

Good luck!!!

Post your picture first ;)

Thanks all!

Dear Mirza,

what is size of you probe, what is temperature of hybridization? Which genomic DNA did you cut? Do you have restriction map of your geneomic dna? please provide these and also your hybridization results, we will give good interpretation? good luck?????