Methods in Molecular Biology • 16 Enzymes of Molecular Biology

BAL 31 Nucleases 233 increasing enzyme concentration (9). In the absence ofDNA polymerase- mediated repair, ligation of partiall ...

234 Gray and Lu 1.4. Uses in Manipulation of Nucleic Acids The duplex exonuclease activity of the nucleases has been very widely ...

BAL 31 Nucleases 235 Enzyme Requirements 2.1. pH The near-neutral pH optima for the nuclease-catalyzed reactions (4,9,25) is an ...

236 Gray and Lu pairs. Moreover, the half-life of the activity on single-stranded DNA and the duplex exonuclease activity are 3- ...

BAL 31 Nucleases 237 single- and double-stranded substrates in the presence of 5% SDS if both Mg 2÷ and Ca 2÷ were present prior ...

238 Gray and Lu Systematic studies have been done on the effects of a "chaotropic" agent (NaC104) and compounds that protect cel ...

BAL 31 Nucleases 239 the huclease with increasing extent of exposure to protease (7). This indi- cates a resistance of the break ...

240 Gray and Lu exonucleolytic attack from the 5' ends, indicate that 90% of Vmax app for the S and F nucleases (10) can be obta ...

BAL 31 Nucleases 241 by overnight stirring. NaOH (2M or higher) is added to a nominal concentration of 0.1M and, after several m ...

242 Gray and Lu where Vto t is the total volume of the acidified reaction mixturealiquot in mL (Vto t = 1.24 mL if the above vol ...

BAL 31 Nucleases 243 corresponding to enzyme nonspecifically bound to duplex DNA away from the ends and not catalytically active ...

244 Gray and Lu in an ordinary agarose gel electrophoresis experiment. For duplex DNA of this G + C content, the value of v0/[S] ...

BAL 31 Nucleases 245 nuclease/L and 58 + 2 nM for gm app. Vma app in Eq. (2) is obtained simply by multiplying the normalized ma ...

246 Gray and Lu with S 1 nuclease or DNA polymerase to remove single-stranded ends and provide fully based-paired termini is unn ...

BAL 31 Nucleases 247 sites was analyzed in this laboratory for both the S and F nucleases, this tendency was confirmed for the S ...

248 Gray and Lu Whether the modifying treatment itself introduces strand breaks is readily ascertained by electrophoresis of the ...

BAL 31 Nucleases 249 Acknowledgments Work reported here for which support has not been acknowledged elsewhere was supported in p ...

250 Gray and Lu ity as a probe for regions of altered secondary structure in negatively and posi- tively supercoiled closed circ ...

BAL 31 Nucleases 251 erties and mechanism of exonuclease action of BAL 31 nuclease. Ph.D. disser- tation, University of Houston. ...

CHAPTER 12 Mung-Bean Nuclease I (EC 3.1.30.1) Andrew J. Sharp and Robert J. Slater Introduction Mung-bean nuclease 1 was first ...