By Kaplan N.P. (ed.), Colowick N.P. (ed.), Wu R. (ed.)
The looks of one other quantity in that very good sequence, equipment in Enzymology, is often a reason for appreciation when you desire to effectively perform a specific procedure or arrange an enzyme or metabolic intermediate with no the tiresome prospect of looking through surprising literature and maybe choosing an unproven process which isn't simply reproduced.
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Additional info for Recombinant DNA Part D
Example text
FIG. 3. Restriction map of E. coli-S, cerevisiae shuttle vector pMHI58. Only unique restriction sites are indicated. Abbreviations: as in Fig. 1; ori 2it, the origin of replication of the yeast 2/x plasmid: LEU2, the yeast L E U 2 gene. 48 VECTORSFOR CLONINGDNA [3] site bank ofpJRD158 by that ofpJRD184 would have made relatively little difference due to duplication of preexisting sites, and was therefore not attempted. In the case of YEp7 derivative pJO158, a considerable improvement could easily be achieved simply by replacing the BamHIXhoI region (containing the restriction site bank) by that of pJRDI84.
Sterilize by autoclaving for 20 min at 15 lb RNase A: 10 mg/ml of pancreatic ribonuclease A (BDH) in TE. 3) and 50% (v/v) glycerol Sequencing and hybridization primers: For sequence aoalysis and for the preparation of hybridization probes two deoxynucleotide primers are used. 5'-dG-T-A-A-A-A-C-G-A-C-G-G-C-C-A-G-T-G-3' (master primer) 5'-dA-A-C-A-G-C-T-A-T-G-A-C-C-A-T-3' (reverse primer) They are complementary to the I and F strands of pKUN, respectively (see Fig. 2B). Primers may be obtained from one of the following companies: Pharmacia, P-L Biochemicals, Bethesda Research Laboratories, Amersham, or Promega Biotec.
4 kb, 9 unique sites48). pJRD215 retains its wide-host-range properties and can be transferred to many different species of gram-negative bacteria in the presence of a self-transmissible plasmid in the donor strain. Conjugation is an efficient process and this may facilitate transfer to species which carry hostile restriction systems. In this context it is interesting to speculate that the statistically abnormal scarcity of 6-bp palindromic restriction sites in 46 R. Meyer, M. Hinds, and M. Brasch, J.