Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 15
The product of the self - primed synthesis of the second strand was a double -
stranded cDNA molecule closed at the terminus corresponding to the 5 ' terminus
of the mRNA by a hairpin loop . This loop was then digested with the single ...
The product of the self - primed synthesis of the second strand was a double -
stranded cDNA molecule closed at the terminus corresponding to the 5 ' terminus
of the mRNA by a hairpin loop . This loop was then digested with the single ...
Page 64
Protocol for the Synthesis of the Second Strand of cDNA The unlabeled first -
strand reaction needs no further treatment before proceeding to the synthesis of
the second strand of cDNA . 1 . Add the following reagents directly to the first -
strand ...
Protocol for the Synthesis of the Second Strand of cDNA The unlabeled first -
strand reaction needs no further treatment before proceeding to the synthesis of
the second strand of cDNA . 1 . Add the following reagents directly to the first -
strand ...
Page 65
second - strand molecules have been generated by self - priming rather than by
replacement synthesis . This can be verified as follows : a . Treat two aliquots of
radiolabeled first strand and two aliquots of radiolabeled second strand with 1
mm ...
second - strand molecules have been generated by self - priming rather than by
replacement synthesis . This can be verified as follows : a . Treat two aliquots of
radiolabeled first strand and two aliquots of radiolabeled second strand with 1
mm ...
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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