Athurva Gore/LabNotes/2011-4-8: Difference between revisions
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*** Heat in formamide or basic solution at 95 for 2 min or 65 for 5 min | *** Heat in formamide or basic solution at 95 for 2 min or 65 for 5 min | ||
*** Boil beads | *** Boil beads | ||
* We can skip desulfonation step according to a published paper: [http://nar.oxfordjournals.org/content/35/1/e4.full?searchid=1&HITS=10&hits=10&resourcetype=HWCIT&maxtoshow=&RESULTFORMAT=&FIRSTINDEX=0&fulltext=tetzner] | |||
** Can use 30 min at 95 C in initial reaction |
Revision as of 22:29, 8 April 2011
Bisulfite Biotin-Binding
- Ran gel for Bisulfite-converted bead-bound DNA and normal bead-bound DNA:
File:04072011 Low Input Bisulfite FAILED.jpg
- Bisulfite-converted libraries did not work.
- DNA was definitely attached to beads, as normal amplification worked.
- DNA must have been detached
- Can we modify the experiment to still allow attachment to beads?
- According to Invitrogen website [1], bead-detachment can occur when:
- Heat in formamide or basic solution at 95 for 2 min or 65 for 5 min
- Boil beads
- According to Invitrogen website [1], bead-detachment can occur when:
- We can skip desulfonation step according to a published paper: [2]
- Can use 30 min at 95 C in initial reaction