Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis
Volume 240, Issue 4, 29 July 1971, Pages 522-531
A crude nuclease preparation suitable for use in DNA reassociation experiments
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Diversity visualization by endonuclease: A rapid assay to monitor diverse nucleotide libraries
2011, Analytical BiochemistryCitation Excerpt :We found that half of the S1 reaction buffer concentration (100 mM NaCH3COO, 0.75 M NaCl, and 5 mM ZnSO4) is sufficient for our assay because the high concentration of salt present in the buffer retarded band migration during electrophoresis. The final DNA concentration used was fixed at 200 ng in 15 μl (13.3 μg/ml) and, hence, was well above the reported concentration of less than 5 μg/ml denatured DNA at which the S1 nuclease reaction results in uncharacteristic reaction kinetics [20]. The variation in band intensity of the 4IE3 sample (Fig. 2B) showed that an excess amount of enzyme results in nonspecific degradation of denatured and reannealed dsDNA.
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Copyright © 1971 Published by Elsevier B.V.