Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-32
... transcription of linearized plasmid DNA contain- ing a bacteriophage promoter are : • 40 mM Tris Cl ( pH 7.5 ) 6 mM MgCl2 2 mm spermidine HCl 5 mM NaCI 10 mm dithiothreitol 100 μg / ml bovine serum albumin ( Fraction V ; Sigma ) 20 nm ...
... transcription of linearized plasmid DNA contain- ing a bacteriophage promoter are : • 40 mM Tris Cl ( pH 7.5 ) 6 mM MgCl2 2 mm spermidine HCl 5 mM NaCI 10 mm dithiothreitol 100 μg / ml bovine serum albumin ( Fraction V ; Sigma ) 20 nm ...
Page 10-36
... transcription systems include : • No RNA synthesis . The most common cause of an apparent lack of RNA synthesis is contamination of tubes or reagents with RNAase . This can be avoided by taking the precautions described in Chapter 7 ...
... transcription systems include : • No RNA synthesis . The most common cause of an apparent lack of RNA synthesis is contamination of tubes or reagents with RNAase . This can be avoided by taking the precautions described in Chapter 7 ...
Page 14-20
A Laboratory Manual Joseph Sambrook, Tom Maniatis. Amplification of DNA Generated by Reverse Transcription of mRNA The first strand of cDNA generated by reverse transcription of mRNA can be used as a template for polymerase chain ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. Amplification of DNA Generated by Reverse Transcription of mRNA The first strand of cDNA generated by reverse transcription of mRNA can be used as a template for polymerase chain ...
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Common terms and phrases
agarose gel aliquots amplification annealing antibody bacterial bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase concentration containing deletions denatured digestion dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Incubate inserted Klenow fragment ligation linear linkers method microfuge tube minutes at 4°C molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probes purified radioactivity radiolabeled reaction mixture recombinant remove restriction enzyme reverse transcriptase room temperature samples screening sequencing reactions single-stranded DNA solution step Store strand of cDNA synthesis target DNA target sequence template DNA termini transfer Tris Cl pH vector