In-solution SureSelect capture library to prepped sample ratio

JUdw · 07-23-2010, 06:20 AM

I’m wondering about the in-solution SureSelect library to prepped sample ratio. The SureSelect protocol states “The ratio of SureSelect capture library to prepped library is critical for successful capture”. What ratio we’re talking about here? I suppose it is the molar bait to target ratio? So then when I have a 4x tilled bait design, can I decrease the Sureselect capture library weight by two? Furthermore, when I enrich a species like e.g. Arabidopsis, which is >10x smaller then human, then the molar bait to target ratio also increases doesn’t it?

Exeplex · 08-18-2010, 08:35 AM

Hi,

There is some useful information in the MYselect manual (a system not dissimilar to Agilent's SureSelect), where they actually give you such details as the bait concentration and numbers of molecules in the bait library versus target (details notably absent from any Agilent protocols).

Should be particularly useful if you are scaling your reactions up or down, doing multiplex capture reactions or using custom libraries.

The MYselect protocol manual can be downloaded from here:
http://www.mycroarray.com/products/myselect.html
and no, I don't work for them (although I'll probably be trying out their system for myself pretty soon)!

Good luck!

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07-23-2010, 06:20 AM	#1
JUdw Junior Member Location: The Netherlands Join Date: Dec 2009 Posts: 9	In-solution SureSelect capture library to prepped sample ratio I’m wondering about the in-solution SureSelect library to prepped sample ratio. The SureSelect protocol states “The ratio of SureSelect capture library to prepped library is critical for successful capture”. What ratio we’re talking about here? I suppose it is the molar bait to target ratio? So then when I have a 4x tilled bait design, can I decrease the Sureselect capture library weight by two? Furthermore, when I enrich a species like e.g. Arabidopsis, which is >10x smaller then human, then the molar bait to target ratio also increases doesn’t it?

08-18-2010, 08:35 AM	#2
Exeplex Member Location: Exeter, UK Join Date: Jun 2009 Posts: 10	Hi, There is some useful information in the MYselect manual (a system not dissimilar to Agilent's SureSelect), where they actually give you such details as the bait concentration and numbers of molecules in the bait library versus target (details notably absent from any Agilent protocols). Should be particularly useful if you are scaling your reactions up or down, doing multiplex capture reactions or using custom libraries. The MYselect protocol manual can be downloaded from here: http://www.mycroarray.com/products/myselect.html and no, I don't work for them (although I'll probably be trying out their system for myself pretty soon)! Good luck!