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Anorexia anx/anx mutant mice (view source)
Revision as of 12:06, 29 November 2024
, 12:06, 29 November 2024→Digestion: added digestion recipe
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===Digestion===
===Digestion===
+
+* 1 ul [https://www.neb.com/en-us/products/r0126-nlaiv NlaIV] (2 units)
+* 5 ul [https://www.neb.com/en-us/products/restriction-endonucleases/hf-nicking-master-mix-time-saver-other/restriction-endonucleases/cutsmart rCutSMart Buffer 10x]
+* 1 ug clean Tyro 3 PCR product
+* H<sub>2</sub>O to 50 ul
+
+Incubate at 37 C for 1 hour, heat inactivate 65 C for 20 min.
NEBiolabs says to mix digest with purple dye with 0.5% SDS to separate protein from DNA. SDS may interfere with SYBR Safe, so may need to post-stain the gel.
NEBiolabs says to mix digest with purple dye with 0.5% SDS to separate protein from DNA. SDS may interfere with SYBR Safe, so may need to post-stain the gel.