Matt:LabNotes/2013-6-27
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Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-6-26
Ethanol Precipitation after PAGE size selection[edit]
- Spun 8 tubes in -80C at 10,000rpm at 4C for 30min
- Discarded supernatant and added 650ul 75% EtOH to wash and spin at 14,000rpm at 4C for 5 min
- Discarded supernatant and let dry in hood 10 min
- Resuspended each of the 8 tubes with 10ul H2O (80ul total)
TBU Gel Quantification Plan[edit]
- Dilute 2ul into 20ul
- Mix 1:1 with 2X buffer (20X dilution)
- Load sample wells with 4,6,8,10,12 ul
- Load Low Mass DNA ladders in 0.5, 1, and 2 ul
' | 0.5 Ladder | 1 Ladder | 4 ul | 6 ul | 8 ul | 10 ul | 12 ul | 2 Ladder |
DNA volume | 0.5 | 1 | 0.2 | 0.3 | 0.4 | 0.5 | 0.6 | 2 |
H2O | 1.5 | 1 | 1.8 | 2.7 | 3.6 | 4.5 | 5.4 | 0 |
2X Buffer | 2 | 2 | 2 | 3 | 4 | 5 | 6 | 2 |
Total | 4 | 4 | 4 | 6 | 8 | 10 | 12 | 4 |
File:2013-06-27 0-gap-probes GelQuantification.jpg
- 20X diluated sample concentration is 1.42 ng/ul
- Starting sample concentration: 28.4 ng/ul -> total yield about 2.2ug/(193nt*325 Da/nt) = 35.1 pmole
Continued on: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-6-28