Streptavidin bead buffers for mate pair prep kit

[slockton]

Hello all,

I have prematurely run out of the three buffers that come with Illumina's Mate Pair Library Prep Kit. Specifically, they are:

• Streptavidin Bead Binding Buffer - part # 1006643
• Streptavidin Bead Wash Buffer 1 - part # 1006644
• Streptavidin Bead Wash Buffer 2 - part # 1006645

Does anyone know what these buffers are and have the recipes for them? I doubt very much you can order them separately from Illumina. Any help would be much appreciated.

All the best,
Steve

[ScottC]

Have you tried asking illumina tech support?

I don't have recipes for these exact buffers, but you can find the recipes for other companies' streptavidin magnetic bead washing buffers on the web. Often they're just PBS pH 7.2 with 0.01% tween-20... but often they're using bound antibodies rather than DNA.

[Bruce E]

Illumina Mate Pair Prep

Did you ever find out what these were. My Mate pair kit is on backorder and I am trying to by reagents separately. I am also trying to which DNA Exonuclease they use.

[slockton]

I eventually got the dynabeads kilobase binder kit from invitrogen since I was doing long insert preps. I substituted the illumina binding protocol for the invitrogen on at the binding stages. I'm not sure this is what you're looking for, though.

[talk24]

I really need to know what exonuclease is used following circularization. I used up all mine experimenting with this step and don't want to choose the wrong one. Anyone know an appropriate exo to use?

[slockton]

I talked to someone from NEB about digesting linear DNA only (and not circularized DNA), and they recommended Lambda Exonuclease (M0262S).

[Paulo.jt]

Does anybody know about the buffers?
What´s the difference between the wash buffers 1 and 2?