Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 12-21
A Laboratory Manual Joseph Sambrook, Tom Maniatis. SCREENING BACTERIAL COLONIES Preparation of Colonies for Screening METHOD 1 This procedure ( Hanahan and Meselson 1980 ) is used when you know in advance that you will need to screen ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. SCREENING BACTERIAL COLONIES Preparation of Colonies for Screening METHOD 1 This procedure ( Hanahan and Meselson 1980 ) is used when you know in advance that you will need to screen ...
Page 12-22
... colonies to regenerate . Generally , it is best to make only two replicas from a single master to avoid problems caused by smearing of the colonies . 10. To induce the expression of a gene cloned into a plasmid carrying the lac promoter ...
... colonies to regenerate . Generally , it is best to make only two replicas from a single master to avoid problems caused by smearing of the colonies . 10. To induce the expression of a gene cloned into a plasmid carrying the lac promoter ...
Page 15-72
... colonies almost certainly contain a mixture of wild - type and mutated molecules , it is necessary to carry out a second round of transformation and screening as outlined below . Note that simple restreaking of positive colonies does ...
... colonies almost certainly contain a mixture of wild - type and mutated molecules , it is necessary to carry out a second round of transformation and screening as outlined below . Note that simple restreaking of positive colonies does ...
Contents
SYNTHESIS OF THE FIRST STRAND OF cDNA 8 | 8-11 |
MOLECULAR CLONING OF DOUBLESTRANDED cDNA 8 | 8-21 |
8 | 8-29 |
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Common terms and phrases
acrylamide agarose gel aliquots amplification antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter chromatography cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing ddNTPs deletions denatured described digestion diluted dithiothreitol DNA fragments DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers method MgCl2 microfuge tube minutes at 4°C molecules mRNA mutants Natl nitrocellulose nitrocellulose filters nucleic acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing reactions solution specific activity step stored strand of cDNA synthesis target DNA target sequence template DNA termini transcription transfer Tris Cl pH vector