Matt:LabNotes/2013-6-27: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>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
 
(3 intermediate revisions by the same user not shown)
Line 10: Line 10:
*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 2, 4, 6, 8 ul
*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 ladder'''
| align="center" style="background:#f0f0f0;"|'''0.5 Ladder'''
| align="center" style="background:#f0f0f0;"|'''1 ladder'''
| align="center" style="background:#f0f0f0;"|'''1 Ladder'''
| align="center" style="background:#f0f0f0;"|''' 2 ul '''
| align="center" style="background:#f0f0f0;"|'''4 ul'''
| align="center" style="background:#f0f0f0;"|''' 4 ul '''
| align="center" style="background:#f0f0f0;"|'''6 ul'''
| align="center" style="background:#f0f0f0;"|''' 6 ul '''
| align="center" style="background:#f0f0f0;"|'''8 ul'''
| align="center" style="background:#f0f0f0;"|''' 8 ul '''
| align="center" style="background:#f0f0f0;"|'''10 ul'''
| align="center" style="background:#f0f0f0;"|'''2 ladder'''
| align="center" style="background:#f0f0f0;"|'''12 ul'''
| align="center" style="background:#f0f0f0;"|'''2 Ladder'''
|-
|-
| DNA volume||0.5||1||0.1||0.2||0.3||0.4||2
| DNA volume||0.5||1||0.2||0.3||0.4||0.5||0.6||2
|-
|-
| H2O||1.5||1||0.9||1.8||2.7||3.6||0
| H2O||1.5||1||1.8||2.7||3.6||4.5||5.4||0
|-
|-
| 2X Buffer||2||2||1||2||3||4||2
| 2X Buffer||2||2||2||3||4||5||6||2
|-
|-
| Total||4||4||2||4||6||8||4
| Total||4||4||4||6||8||10||12||4
|-
|-
|  
|  
|}
|}


*20X diluated sample concentration is 1ng/ul
[[File:2013-06-27_0-gap-probes_GelQuantification.jpg | 650px]]
*Starting sample concentration: 20ng/ul -> total yield about 1.2ug/(193nt*325 Da/nt) = 19.1 pmole
 
*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