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=Testing "Homemade" Reagents to be used with CircLigase II= *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== *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== [[File:20151007_CircLigaseIIReagentsTest.jpg|650px]] *Hosuk's buffer works well *5M Betaine by Justin does not work **Use 5M Betaine from SigmaAldrich *50mM MnCl2 works decently
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