used were 0 1 tfa in water a 70 acetonitrile in water 0 070 tfa b flow rate was 150 u03bcl per minute with a gradient as follows 0 minutes u2014 5 buffer b 10 minutes u2014 10 buffer b 80 minutes u2014 80 buffer b 85 minutes u2014 100 buffer b 90 minutes u2014 100 buffer b cnbr digests are treated as follows water is added to the solution and the whole volume concentrated to 100 u03bcl in a speed vac further water is added to approximately 1 ml and this dried again to about 100 u03bcl this removes the majority of the formic cnbr various peptide fragments obtained as described above were analyzed to determine their sequence and for subsequent development of degenerate probes for use in cloning the gene encoding the 38 kd esterase from the genome of the donor organism peptide sequence analysis of the 38 kd esterase was problematic due to cycles containing mixed signals indicating the presence of multiple polypeptides in the analyzed sample protein sequencing resulted in an n terminal sequence and several additional peptide fragments as follows from these fragments suitable degenerate probes for hybridization and use as pcr primers were produced and fragments were obtained which were believed to be derived from the gene encoding the 38 kd esterase however sequencing of the fragments obtained in this manner 550 and 100 base pairs showed that the fragments were merely artifacts of pcr and were not of use in cloning the 38 kd esterase additional analysis of 2 different probes derived from 2 protein sequences isolated as above resulted in similar lack of success from these results it was determined that routine protein purification and peptide.