AlanFung:LabNotes/CTCF/2011-8-26: Difference between revisions

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>Alan6017518
(Created page with '===Make New BSA-Water solution=== *Stock solution is at 10mg/ml, need to dilute to 0.5mg/ml *Take 10ul stock BSA dilute with 190ul nuclease free water ===Bisulfite Conversion - I…')
 
>Alan6017518
Line 4: Line 4:
===Bisulfite Conversion - Imprint Kit===
===Bisulfite Conversion - Imprint Kit===
* Prepare DNA Modification Solution (CT Conversion Reagent):  
* Prepare DNA Modification Solution (CT Conversion Reagent):  
* Add 21 uL 0.5 mg/mL BSA and 1 uL Imprint Balance solution to each sample; incubate at 37 C for 10 minutes
* Add 21 uL 0.5 mg/mL BSA and  
*Add1 uL Imprint Balance solution to each sample;  
*incubate at 37 C for 10 minutes
* Add 125 uL DNA Modification solution to each tube, incubate at 65 C for 90 minutes
* Add 125 uL DNA Modification solution to each tube, incubate at 65 C for 90 minutes
* Perform Column Purification of each sample:
* Perform Column Purification of each sample:
** Elute into 11uL Elution Buffer
** Elute into 11uL Elution Buffer
*Denature all samples at 95C for 30sec and quickly transfer to cooler rack
*Denature all samples at 95C for 30sec and quickly transfer to cooler rack

Revision as of 02:52, 27 August 2011

Make New BSA-Water solution

  • Stock solution is at 10mg/ml, need to dilute to 0.5mg/ml
  • Take 10ul stock BSA dilute with 190ul nuclease free water

Bisulfite Conversion - Imprint Kit

  • Prepare DNA Modification Solution (CT Conversion Reagent):
  • Add 21 uL 0.5 mg/mL BSA and
  • Add1 uL Imprint Balance solution to each sample;
  • incubate at 37 C for 10 minutes
  • Add 125 uL DNA Modification solution to each tube, incubate at 65 C for 90 minutes
  • Perform Column Purification of each sample:
    • Elute into 11uL Elution Buffer
  • Denature all samples at 95C for 30sec and quickly transfer to cooler rack