Used gel-purified, size-selected DNA from yesterday to prepare the RAD library using the Kappa LTP Kit:

http://eagle.fish.washington.edu/trilobite/Sites_genefish_100112/Steven/Commercial%20Protocols/KAPA_Biosystems%20-%20KAPA_LTP_Library_Preparation_Kit_TDS.pdf

The protocol was followed with the following changes:

– Section 8

Skipped entirely

– Section 9.1

Used 10uL of library DNA (instead of 20uL)

Used 1uL of mixed primer set (instead of 5uL)

– Section 9.2

Performed 12 cycles of PCR protocol. This was Carita’s recommendation and experience with using the Kappa LTP Kit for RAD library construction.

Sample was eluted from the AMPure beads with 15uL of Buffer EB (Qiagen) and stored @ -20C.