Reduced Representation library protocol

[fastidiousme]

Could someone kindly tell me the procedure of selecting one restriction enzyme for an entire genome, for the purpose of constructing a reduced representation library?

[protist]

Reduced Represenation Library Protocol

Hi Helena,

Check out a recent Nature Protocols, while it is directed towards methylation sites utilising a methylation-insensitive restriction enzyme MspI it should give you some pointers. It is very descriptive and has nice how-to gel pictures. Best of luck.

http://www.nature.com/nprot/journal/v6/n4/full/nprot.2010.190.html

[Rob_Elshire]

Our Paper is out

Hi,

You can find the paper about the above work here:
A robust, simple genotyping-by-sequencing (GBS) approach for high diversity species.

Best regards,

Rob Elshire

[luc]

This site might be helpful:

http://www.maizegenetics.net/images/stories/GBS/Buckler_Lab_Genotyping_by_Sequencing_Protocol_20110114.pdf

http://www.maizegenetics.net/Table/Genotyping-By-Sequencing/

[[email protected]]

Hi All,

I am going to prep some RRL to run on the solexa GAIIx and was wondering if anyone had working protocol they could share. I have ordered the enzymes and have read most of the recent publications but the methods are brief.

The main details I would like to clarify are:
Volume of starting DNA
Timing of cleanup steps
Is there a need to run two gel through the protocol, per and post adaptor ligation?

Thank you in advance for your help.