Matt:LabNotes/2013-6-27: Difference between revisions
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>Mzcai (Created page with "Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-6-26 ===Ethanol Precipitation after PAGE size selection=== *Spun 8 tubes in -80C at 10,000rp...") |
>Mzcai mNo edit summary |
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*Dilute 2ul into 20ul | *Dilute 2ul into 20ul | ||
*Mix 1:1 with 2X buffer (20X dilution) | *Mix 1:1 with 2X buffer (20X dilution) | ||
*Load sample wells with | *Load sample wells with 4,6,8,10,12 ul | ||
*Load Low Mass DNA ladders in 0.5, 1, and 2 ul | *Load Low Mass DNA ladders in 0.5, 1, and 2 ul | ||
{| {{table}} | {| {{table}} | ||
| align="center" style="background:#f0f0f0;"|'''''' | | align="center" style="background:#f0f0f0;"|'''''' | ||
| align="center" style="background:#f0f0f0;"|'''0.5 | | align="center" style="background:#f0f0f0;"|'''0.5 Ladder''' | ||
| align="center" style="background:#f0f0f0;"|'''1 | | align="center" style="background:#f0f0f0;"|'''1 Ladder''' | ||
| align="center" style="background:#f0f0f0;"|''' | | align="center" style="background:#f0f0f0;"|'''4 ul''' | ||
| align="center" style="background:#f0f0f0;"|''' | | align="center" style="background:#f0f0f0;"|'''6 ul''' | ||
| align="center" style="background:#f0f0f0;"|''' | | align="center" style="background:#f0f0f0;"|'''8 ul''' | ||
| align="center" style="background:#f0f0f0;"|''' | | align="center" style="background:#f0f0f0;"|'''10 ul''' | ||
| align="center" style="background:#f0f0f0;"|'''2 | | align="center" style="background:#f0f0f0;"|'''12 ul''' | ||
| align="center" style="background:#f0f0f0;"|'''2 Ladder''' | |||
|- | |- | ||
| DNA volume||0.5||1||0. | | DNA volume||0.5||1||0.2||0.3||0.4||0.5||0.6||2 | ||
|- | |- | ||
| H2O||1.5||1 | | H2O||1.5||1||1.8||2.7||3.6||4.5||5.4||0 | ||
|- | |- | ||
| 2X Buffer||2||2 | | 2X Buffer||2||2||2||3||4||5||6||2 | ||
|- | |- | ||
| Total||4||4 | | Total||4||4||4||6||8||10||12||4 | ||
|- | |- | ||
| | | | ||
|} | |} | ||
*20X diluated sample concentration is | [[File:2013-06-27_0-gap-probes_GelQuantification.jpg | 650px]] | ||
*Starting sample concentration: | |||
*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 |
Latest revision as of 18:41, 28 June 2013
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