Matt:JustinLabNotes/2015-10-7
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Testing "Homemade" Reagents to be used with CircLigase II[edit]
- CircLigase II kit does not provide enough 5M Betaine, 50mM MnCl2, and 10X CircLigaseII Buffer for DARTFISH volumes
- Test 10X Buffer Hosuk made
- Test 5M Betaine Justin made from dry (seems very old)
- Test 50mM MnCl2 Justin made from Manganese chloride tetrahydrate (also seems very old from Paola's lab)
Protocol[edit]
- Followed protocol provided by CircLigase II kit and control ssDNA in kit
- Try substituting one "homemade" reagent for kit provided reagent in each sample
Results[edit]
File:20151007 CircLigaseIIReagentsTest.jpg
- Hosuk's buffer works well
- 5M Betaine by Justin does not work
- Use 5M Betaine from SigmaAldrich
- 50mM MnCl2 works decently