amount of DNA for clontech genome walker system - how much (Apr/25/2005 )
thanks to without00.
anyway, i am interested in clontech's GenomeWalker System. but it's said it needs 2-5ug genomic DNA, is that true? why we need so much? can I go ahead with about hundreds of ng DNA? it is for plant(arabidopsis).
thank you for your reply.
You must mean the universal genomewalker kit. It may need 2-5 ug DNA to build four libraries with 4 restriction enzyme. After you have built the libraries, they are good for quite a few PCRs.
Thanks a lot.
I have another kind of trivial problem for this kit.
Since we don't have TE buffer of Ph7.5 but Ph 8.5(at 25C). Can we just use this or pure water instead to dissolve the digested DNA? To put it the other way, does the PH value of the buffer severly affect the following ligation step inspite of the ligation buffer? Also,since the Ph value of the Tris would decrease(0.03-0.05 per 10 degrees) during the ligation (16C), then shouldn't we use Ph8 instead of 7.5 giving the optimal Ph for ligation is 7.2-7.8?
Any one can give me some explanation?