Arichard:Notebook/bulk150122: Difference between revisions

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>Andrew
(Created page with "==Bulk libraries January 22, 2015== [http://genome-tech.ucsd.edu/LabNotes/index.php/Brandon:LabNotes/Project1/2014-11-18 Brandon's protocol for DNA purification after guanidi...")
 
>Andrew
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[http://genome-tech.ucsd.edu/LabNotes/index.php/Brandon:LabNotes/Project1/2014-11-18 Brandon's protocol for DNA purification after guanidine HCl]
[http://genome-tech.ucsd.edu/LabNotes/index.php/Brandon:LabNotes/Project1/2014-11-18 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.
ADD MORE METHODS
Ran first gel after first Ampure. 3 out of 20 ul of each sample loaded.
[[File:bulk150122Slide1.jpg|400px]]
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|400px]]
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|400px]]
2X Ampure XP worked well. Smears look equivalent to PA size selection. Samples 1 and 2 submitted (QJ01 and QJ02)

Revision as of 00:38, 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.

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)