Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 22
Heat the mixture to 85°C for 5 minutes and then let it cool slowly to 37°C . This
can be achieved by floating the tube in a piece of Styrofoam in a 250 - ml beaker
filled with water equilibrated to 85°C . Store the beaker at room temperature until
...
Heat the mixture to 85°C for 5 minutes and then let it cool slowly to 37°C . This
can be achieved by floating the tube in a piece of Styrofoam in a 250 - ml beaker
filled with water equilibrated to 85°C . Store the beaker at room temperature until
...
Page 36
Add 5 – 10 volumes of the EDTA - Tris - DNA solution to a microfuge tube
containing the solution of radiolabeled oligonucleotide that is to be precipitated .
Mix well . For precipitation to be efficient , it is important that the ionic composition
of the ...
Add 5 – 10 volumes of the EDTA - Tris - DNA solution to a microfuge tube
containing the solution of radiolabeled oligonucleotide that is to be precipitated .
Mix well . For precipitation to be efficient , it is important that the ionic composition
of the ...
Page 9
Using a fresh pipette tip , transfer 10 ul of the enzyme : DNA mixture to the
second tube . Mix quickly , and replace the tube ... Continue this procedure until
seven of the tubes contain both restriction enzyme and DNA . The eighth tube ,
which ...
Using a fresh pipette tip , transfer 10 ul of the enzyme : DNA mixture to the
second tube . Mix quickly , and replace the tube ... Continue this procedure until
seven of the tubes contain both restriction enzyme and DNA . The eighth tube ,
which ...
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Contents
TEST LIGATION 3 29 | 3 |
SYNTHESIS OF THE FIRST STRAND OF cDNA 8 | 11 |
FRAGMENT OF E coli DNA POLYMERASE I 13 59 | 14 |
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Common terms and phrases
acid activity agarose aliquots allow amount amplification appropriate approximately bacteriophage bacteriophage T4 base buffer carried cDNA cells centrifugation Chapter chromatography cleaved clones colonies concentration constructed containing deletions denatured described determine digestion DNA polymerase dNTPs double-stranded efficiency electrophoresis et al ethanol expression Figure filters fragment gene genomic hybridization increase Incubate inserted interest isolated labeled length libraries ligation linear linkers method microfuge minutes mixture molecules mRNA mutagenesis mutagenic mutations Nucleic nucleotides obtained oligonucleotide original plaques plasmid plasmid DNA plate pool position possible prepared presence primer probe problem protein purified radiolabeled reaction recombinant region remove restriction enzyme room temperature samples screening sequence single single-stranded solution specific step Store strand synthesis target DNA target sequence template termini tion transfer transformants tube units usually vector volume