A modification of the Kunkel method for preparation of single-stranded DNA with helper phage M13 KO7
نویسندگان
چکیده
منابع مشابه
Improved method for the production of M13 phage and single-stranded DNA for DNA sequencing.
An improved method is described for the efficient production of M13 phage and M13 single-stranded (ss)DNA in a relatively short time period. Infection of E. coli (F') cells with as few as 5 phage particles can yield 10(12) phage particles/mL in 3 hours if the cells are grown in LB broth or SOB broth supplemented with about 5 mM Mg2+. The method tolerates large variations in the initial multipli...
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Conformations of the single-stranded DNA of bacteriophage M13.
At least two conformations of M13 single-stranded DNA have been demonstrated by measuring differences in sedimentation coefficient and by direct visualization in the electron microscope. Which form is obtained from infected cells and/or intact phage depends on the pH, ionic strength, and temperature. The slower-sedimenting form can be converted to the faster-sedimenting, single-stranded form by...
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The replication origins of viral and complementary strands of bacteriophage M13 DNA are contained within a 507-nucleotide intergenic region of the viral genome. Chimeric plasmids have been constructed by inserting restriction endonuclease fragments of the M13 intergenic region into the plasmid pBR322. Replication of these hybrid plasmids, under conditions not permissive for the plasmid replicon...
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ژورنال
عنوان ژورنال: Technical Tips Online
سال: 1997
ISSN: 1366-2120
DOI: 10.1016/s1366-2120(08)70031-x