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文献和实验Attachment of Nitroxide Spin Labels to Nucleic Acids for EPR
Couet, W.R., Brasch, R.C., Sosnovsky, C., Lukszo, J., Prakash, I., Gnewech, C.T., and Tozer, T.N. 1985. Influence of chemical structure of nitroxyl spin labels on their reduction by ascorbic acid. Tetrahedron 41:1165‐1172
SOUTHERN BLOTTING OF GENOMIC DNA
1.Digest DNA: • 1.5 μg DNA in a 25 μI reaction containing BSA • 3 to 5 units for 5 to 3 hours (aim for 10-fold overdigestion; remember many enzymes will not survive well for more than 3 hr). • Also prepare molecular weight standard (Lambda
PCR amplification of variable regions (SOE-Protocol)
in the supernatant through a S-400 sephacryl micros-pin column: a. vortex the micro-spin column to homogenize b. screw off the top cap but leave it on c. break off the bottom cap d. put the column in a eppendorf tube and spin for 1 min at 735 xg e. pipet
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