Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-4
... 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 . The following tests are commonly used : • The ability of 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 . The following tests are commonly used : • The ability of the ...
Page 8-6
... mRNA preparations that are rich in the sequences of interest , or of mRNA that has been partially fragmented by limited alkaline hydrolysis and end - labeled by phosphor- ylation . As a good approximation , the mRNA sequences of ...
... mRNA preparations that are rich in the sequences of interest , or of mRNA that has been partially fragmented by limited alkaline hydrolysis and end - labeled by phosphor- ylation . As a good approximation , the mRNA sequences of ...
Page 8-48
... mRNA is depleted of sequences that are present in a second type of mRNA by subtractive hybridization ( Timberlake 1980 ; Zimmerman et al . 1980 ) . Typically , the cDNA is hybridized two or three times in succession to a 20 - fold ...
... mRNA is depleted of sequences that are present in a second type of mRNA by subtractive hybridization ( Timberlake 1980 ; Zimmerman et al . 1980 ) . Typically , the cDNA is hybridized two or three times in succession to a 20 - fold ...
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Common terms and phrases
agarose gel aliquots amino acid amplification bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA Biol buffer carried cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing deletions denatured digestion DNA fragments DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol ethidium bromide exonuclease formamide gel electrophoresis gene genomic DNA H₂O hybridization Incubate the reaction inserted Klenow fragment labeled ligation linkers method microfuge tube minutes at 4°C mismatched molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nucleotides oligonu oligonucleotide phagemid plaques plasmid plasmid DNA plates polymerase chain reaction pool primer probe protein purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples screening sequencing reactions single-stranded DNA solution specific activity Store strand of cDNA synthesis target DNA target fragment target sequence template DNA termini transcription transfer Tris Cl pH vector µg/ml