Ampure magnetic bead purification -- how does it work ?

Christine90 · 11-18-2012, 08:42 AM

Hi I'm new in using NGS especially the GS Junior.
I prepaired a library using self designed primers. After amplification there's a purification step by magnetic beads. I understand the basics but not every reaction is clear to me. My main questions are: Why do they say that its equal using 10µl library or fewer and add some water. Doesn't this change my purification result?
In the patent of this technique the salt concentration is mentioned to influence the results. Why and what does salt do. If I add water, my salt concentration is reduced isn't it? Have to explaine this method in my thesis.
Could you please try to explain the chemical background to me.

Thanking you in anticipation!
Regards Christine90

ECO · 11-18-2012, 09:24 AM

There are plenty of threads about the mechanism...please
search, especially if it's for your thesis!

TonyBrooks · 11-19-2012, 05:16 AM

http://core-genomics.blogspot.co.uk/...eads-work.html

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11-18-2012, 08:42 AM	#1
Christine90 Junior Member Location: Bavaria Join Date: Nov 2012 Posts: 3	Ampure magnetic bead purification -- how does it work ? Hi I'm new in using NGS especially the GS Junior. I prepaired a library using self designed primers. After amplification there's a purification step by magnetic beads. I understand the basics but not every reaction is clear to me. My main questions are: Why do they say that its equal using 10µl library or fewer and add some water. Doesn't this change my purification result? In the patent of this technique the salt concentration is mentioned to influence the results. Why and what does salt do. If I add water, my salt concentration is reduced isn't it? Have to explaine this method in my thesis. Could you please try to explain the chemical background to me. Thanking you in anticipation! Regards Christine90

11-18-2012, 09:24 AM	#2
ECO --Site Admin-- Location: SF Bay Area, CA, USA Join Date: Oct 2007 Posts: 1,358	There are plenty of threads about the mechanism...please search, especially if it's for your thesis!

11-19-2012, 05:16 AM	#3
TonyBrooks Senior Member Location: London Join Date: Jun 2009 Posts: 298	http://core-genomics.blogspot.co.uk/...eads-work.html