Bead dilution buffer

[koala]

Hi everyone,
I'm looking for the composition of bead dilution buffer in the purification step after the recovery of circular DNA in mate pair library (used with XP ampure).

The problem I've found is that this buffer ends in a very short time respect to other supplied material in the dynabeads module of the library.

If anybody knows the composition..... or could it be substituted by something similar???

thanks
chiara

[peggywangping]

yeah, we substituted the beads dilution buffer with nuclease free water in the mate pair library (for 3k library, don't know the result for 10k library), and it works. but i am still worried about that the beads dilution buffer contains PEG and Nacl that will affect the size-selection process and final size of DNA fragment.

[koala]

thanks. I've tried too, but I've lost DNA!!

Do you have 5500 xl SOLiD? how many MP libraries have you product? what about insert size?

[peggywangping]

actually, we don't use the reagent in this kit, we just substituted phenol purification step with the ampure xp purification, save more time than previously old protocol. for the DNA fragment, we used about 7-8K genomic DNA. Comparing the phenol and Qiagen kit purification kit, ampure xp beads can give us very unique DNA fragment according to the agilent results.