MIT7_13f08_lab20_Protocol_Restriction

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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 Restriction digests There are two basic types of restriction digests that you will perform during this course. An analytical restriction digest is done when you need to use the information from the digest to create a map of your DNA or to be sure that you have the correct plasmid. A preparative restriction digest is done when you need to prepare a large amount of particular DNA fragment. The key difference is scale. Generalized protocols for both are described below. REAGENTS Restriction enzymes: Stored in -20°C. The enzymes are very expensive and we all must share the same stock. DO NOT CROSS-CONTAMINATE ENZYMES. ALWAYS KEEP THEM ON ICE. Enzyme buffers and 100x (10 mg/ml) BSA These are supplied by New England BioLabs and are coded into 4 different types of buffer. Refer to the NEB catalogue for the correct buffer for each enzyme.
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