BIO372H5 Lecture Notes - Lecture 2: Alpha Beta Gamma, Aspergillus Nuclease S1, Gastrointestinal Tract

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8 Mar 2014
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S1 nuclease mapping of the 5" end of a transcript. How to find start and stop signals - find mrna and sequence it, then align with reference genome. Gamma is on n and that is transferred in the polynucleotide. Si nuclease - degrade single strand but not double strand. All the enzymes were needed to be made in the old days, so this experiment took so long to finish. But now, you can just order from biotech companies. Description: calf intestinal alkaline phosphatase (ciap) is a phosphomonoesterase that removes 3 and 5 phosphates from dna and rna. Applications: dephosphorylation of 5 -phosphorylated termini of vector dna to prevent self-ligation (1). Performance and quality testing: endodeoxyribonuclease, 3 exodeoxyribonuclease, and ribonuclease assays; dephosphorylation efficiency measured in a transformation assay. Unit definition: one unit hydrolyzes 1 mol of 4-nitrophenyl phosphate in 1 min. at 37 . Unit reaction conditions: 1 m diethanolamine buffer, 10 mm 4-nitrophenyl phosphate,