fragmentation buffer of mRNA-seq

[Nik]

Hi,

Did you ever find out about this?

Nik

[xile]

Hi everyone,

I plan to try mRNA-seq sample preparation with my own reagents. I am glad if somebody give me some information of the components of fragmentation buffer and fragmentation stop buffer in Illumina Kit. After google searching, I found that the components are different from different researcher. For example, UCLA:
200 mM Tris-Acetate, pH 8.1, 500 mM KOAc, 150 mM MgOA)

The fragmentation buffer is made with RNase-free reagents. Tris-containing solutions should not be treated with DEPC; however, once H2O has been DEPC-treated and autoclaved it can be used for making the Tris solution.
Combine the following components to a total volume of 20 mL:
4 mL 1 M Tris acetate pH 8.1 (Trizma Base, PH adjusted with glacial acetic acid)
0.64g MgOAc
0.98g KOAc
Depc-treated H2O to 20 ml
Mix throroughly and filter through a 0.2 uM vacuum filter unit. This reagent should be aliquotted and stored at RT.

ULS:
RNA fragmentation buffer: 100mM ZnCl2 in 100mM Tris-HCl pH7
RNA fragmentation stop buffer: 0.5M EDTA pH8
Or
Ambion fragmentation reagent #8740


Can they use for mRNA-seq? which is the better?