By Annette M. Griffin, Hugh G. Griffin
DNA Sequencing Protocols will give you the data to develop into a sequencing professional. An all-star solid of investigators covers almost all elements, together with advancements in cycle sequencing, sequencing PCR items, sequencing lambda and cosmids, multiplex sequencing, direct blotting electrophoresis, sequencing of chemiluminescence, and automatic sequencing.
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Extra info for DNA Sequencing Protocols
Transfer supernatants to clean tubes making sure the pellet is undisturbed, add 200 uL of PEG/NaCl to each, vortex well, and leave at room temperature for at least 15 min. 6. Spin for 5 min in microfuge to pellet the phage particles. Remove the PEG-containing supernatant with a pipet, respin the tubes for a few seconds and carefully remove all remaming traces of supernatant from around the phage pellet using a drawn-out Pasteur pipet (see Note 5). The phage pellet should be visible at the bottom of the tube.
2 Hanahan D. (1983) Studies on transformation of Escher&ia coli with plasmids J Mol. Blol 166,557-580. 3 Sambrook J , Fritsch E. 84 4 Dagert M and Ehrlich S D (1979) Prolonged incubation m calcium chloride improves the competence of Escherichia colr cells Gene 6,23-28 Ml3 Phage Growth and Single-Strand DNA Preparation 1. Introduction Ml3 phages do not lyse their host, but are released from infected cells as the cells continue to grow and divide. Cells infected with Ml3 have a longer replication cycle than uninfected cells, and as the infection proceeds, areas of slower-growing cells can be visualized as turbid plaques on lawns of E.
In this laboratory, deletions are mainly performed on miniprep DNA to 58 Murphy avoid the time-consuming process of caesium gradient banding. The use of spm-columns to clean up linearized DNA provides excellent substrates for dtgestion. However, the digestion rates of miniprep plasmtd DNA tend to be more variable than when using more purified material. If using mmiprep DNA the plasmid from two overnight lo-mL cultures are pooled m a volume of 100 pL. This provides enough material for stx trials of the digestion rate if 30 & is used per lmearization.