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.
Sample 3 (index 4) negative. Mis-loading suspected. QJ ran second gel to confirm. Another 3 ul loaded (14 ul now remaining).
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
2X Ampure XP worked well. Smears look equivalent to PA size selection. Samples 1 and 2 submitted (QJ01 and QJ02)