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 |
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===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 | * 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