Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 4
Integrity of the mRNA Since the cDNA library cannot be better than the mRNA
from which it is derived , it is important to check the integrity of the preparation of
mRNA before it is used as the template for synthesis of the first strand of cDNA .
Integrity of the mRNA Since the cDNA library cannot be better than the mRNA
from which it is derived , it is important to check the integrity of the preparation of
mRNA before it is used as the template for synthesis of the first strand of cDNA .
Page 6
Abundant mRNAS Initially , cDNA cloning was used to obtain copies of abundant
mRNAs such as those encoding globin ... Consequently , no further purification of
the particular mRNA is required before double - stranded cDNA is synthesized ...
Abundant mRNAS Initially , cDNA cloning was used to obtain copies of abundant
mRNAs such as those encoding globin ... Consequently , no further purification of
the particular mRNA is required before double - stranded cDNA is synthesized ...
Page 48
Subtracted cDNA probes Subtracted cDNA probes are often used to probe cDNA
libraries for clones that correspond to mRNAs that are differentially regulated . A
cDNA probe prepared from one type of mRNA is depleted of sequences that are
...
Subtracted cDNA probes Subtracted cDNA probes are often used to probe cDNA
libraries for clones that correspond to mRNAs that are differentially regulated . A
cDNA probe prepared from one type of mRNA is depleted of sequences that are
...
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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