AlanFung:LabNotes/CTCF/2011-5-10: Difference between revisions

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>Alan6017518
Line 15: Line 15:
===Make Serial Dilution on Control DNA===
===Make Serial Dilution on Control DNA===
*Create 6 STDs
*Create 6 STDs
{| {{table}}
| Sample Name||dsDNA Concentration (pM)
|-
| Std 1||20
|-
| Std 2||2
|-
| Std 3||0.2
|-
| Std 4||0.02
|-
| Std 5||0.002
|-
| Std 6||0.0002
|-
|
|}

Revision as of 22:46, 25 May 2011

Bisulfite Conversion DNA Damage Estimation

  • Perform PCR with CHR8, CHR21 and CHR22
  • These will be our standards

File:ZhangLab 2 2011-05-10 10hr 40min.jpg

  • Need to purify CHR8, CHR21, and CHR22.
    • Qubit Results(ng/ul)
    • Assume amplicon length ~200bp
  • CHR8-3.25ng/ul 26.72nM
  • CHR21-4.39ng/ul 36.09nM
  • CHR22-4.77ng/ul 39.21nM

Determine the efficiency of bisulfite conversion

  • After performing the bisulfite conversion procedures, we should perform real-time PCR of the samples and controls to determine the amount of unconverted gDNA vs. converted human genomic DNA. Base on the difference in the CT values, we can determine the conversion efficiency.


Make Serial Dilution on Control DNA

  • Create 6 STDs
Sample Name dsDNA Concentration (pM)
Std 1 20
Std 2 2
Std 3 0.2
Std 4 0.02
Std 5 0.002
Std 6 0.0002