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上海玉博生物科技有限公司
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文献和实验S1 Nuclease Protection Assay End-label oligo (20-25 mer)4 µl 5X T4 Kinase Buffer5 µl [gamma-32P] ATP (7000 Ci/mmol)10 µl water + oligo (200 ng)1 µl T4 Kinase1. 37 deg C for 1 hour.2. Heat inactivate at 75 deg C for 10 minutes.3. Add 1 µl glycogen
1. Cut DNA with 5'- and 3'- overhangs, gel check, phenol-Sevag extract and NaOAc/EtOH ppt. 2. Prepare one S1 nuclease tube (on ice) for each time point: 15ml S1 Buffer + 0.25U S1/ml (5U). 3. Use 5ml (=0.5mg) digested plasmid (in EB) per time
S1 analysis of yeast mRNA using oligonucleotide probes
on the sequencing gel to observe the quality and size of probe with no S1 treatment. Hybridization Mix: 5 microliters 4 M NaCl 2.5 microliters 1 M PIPES pH 6.9 1 microliter 0.25 M EDTA 1-2 x 105 cpm kinased
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