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MIT7_13f08_lab19_Protocol_QIAquickPCR - MIT OpenCourseWare...

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MIT OpenCourseWare http://ocw.mit.edu 7.13 Experimental Microbial Genetics Fall 2008 For information about citing these materials or our Terms of Use, visit: http://ocw.mit.edu/terms .
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7 . 1 3 F a l l 2 0 0 8 P a g e | 1 QIAquick PCR & Enzyme Purification Kit Protocol This protocol is designed to purify single- or double-stranded 100bp-10kb DNA fragments from PCR and other enzymatic reactions. You should read through the QIAquick Spin Handbook posted at the 7.13 Stellar website for in depth information about this protocol. Important points before starting The yellow color of Buffer PBI indicates a pH 7.5. Add ethanol (96–100%) to Buffer PE before use (see bottle label for volume). All centrifugation steps are carried out at 13,000 rpm (~17,900 x g ) in a conventional table-top microcentrifuge. 1. Add 5 volumes of Buffer PBI to 1 volume of the PCR/Enzyme reaction and mix (it is not necessary to remove mineral oil or kerosene). For example, add 500 µl of Buffer PB to 100 µl PCR sample (not including oil).
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MIT7_13f08_lab19_Protocol_QIAquickPCR - MIT OpenCourseWare...

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