Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 27
SEQUENTIAL ADDITION OF DIFFERENT LINKERS In this method ( Helfman et
al . 1983 ) ( Figure 8 . 10 ) , double - stranded cDNA synthesized by the self -
priming mechanism is ligated to one type of synthetic linker before the hairpin
loop in ...
SEQUENTIAL ADDITION OF DIFFERENT LINKERS In this method ( Helfman et
al . 1983 ) ( Figure 8 . 10 ) , double - stranded cDNA synthesized by the self -
priming mechanism is ligated to one type of synthetic linker before the hairpin
loop in ...
Page 7
Enzymes that generate blunt ends ( FnuDII , Alul , Rsal , and HaeIII ) are
particularly useful because they allow the direct addition of a large variety of
synthetic linkers . However , enzymes that cleave in a staggered fashion can be
used if the ...
Enzymes that generate blunt ends ( FnuDII , Alul , Rsal , and HaeIII ) are
particularly useful because they allow the direct addition of a large variety of
synthetic linkers . However , enzymes that cleave in a staggered fashion can be
used if the ...
Page 86
Designing the Hexameric Linker Before starting hexameric linker mutagenesis , it
is necessary to decide ( 1 ) which ... The amino acids that will be inserted in the
mutant protein depend on the reading frame and on the sequence of the linkers .
Designing the Hexameric Linker Before starting hexameric linker mutagenesis , it
is necessary to decide ( 1 ) which ... The amino acids that will be inserted in the
mutant protein depend on the reading frame and on the sequence of the linkers .
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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 |
Copyright | |
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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