Arichard:Notebook/bulk150122: Difference between revisions

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Qiming Jin (QJ) ran most of these samples with my supervision.
Qiming Jin (QJ) ran most of these samples with my supervision.
Used SW480 2N Full sorted October 21, 2014 (tubes labeled 4 in sort box)


ADD MORE METHODS
ADD MORE METHODS

Revision as of 23:22, 4 February 2015

Bulk libraries January 22, 2015

Brandon's protocol for DNA purification after guanidine HCl

Used Blue's method of 2x Ampure XP at 0.8:1 ratio to select 200-800 bp.

Qiming Jin (QJ) ran most of these samples with my supervision.

Used SW480 2N Full sorted October 21, 2014 (tubes labeled 4 in sort box)

ADD MORE METHODS

Ran first gel after first Ampure. 3 out of 20 ul of each sample loaded.

File:Bulk150122Slide1.jpg

Sample 3 (index 4) negative. Mis-loading suspected. QJ ran second gel to confirm. Another 3 ul loaded (14 ul now remaining).

File:Bulk150122Slide2.jpg

Second gel confirms that sample 3 did not amplify in PCR. Performed second Ampure XP purification:

  • 14 ul sample (20 - 3 - 3)
  • 11.2 ul beads
  • Elute in 20 ul H2O

File:Bulk150122Slide3.jpg

2X Ampure XP worked well. Smears look equivalent to PA size selection. Samples 1 and 2 submitted (QJ01 and QJ02)