Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 9-47
... hybridization Although the choice depends to a large extent on personal preference , we offer the following guidelines for choosing among the various methods available . 1. Hybridization reactions in 50 % formamide at 42 ° C are less ...
... hybridization Although the choice depends to a large extent on personal preference , we offer the following guidelines for choosing among the various methods available . 1. Hybridization reactions in 50 % formamide at 42 ° C are less ...
Page 9-48
... hybridization are un- known . When using probes that have the capacity to self - anneal ( e.g. , nick - translated double - stranded DNA ) , the following rule of thumb is useful : Allow the hybridization to proceed for a time ...
... hybridization are un- known . When using probes that have the capacity to self - anneal ( e.g. , nick - translated double - stranded DNA ) , the following rule of thumb is useful : Allow the hybridization to proceed for a time ...
Page 9-50
... hybridization signal is particularly noticeable when oligonucleotides or probes less than 100 nucleotides in length are used . 7. In the presence of 10 % dextran sulfate or 10 % polyethylene glycol , the rate of hybridization is ...
... hybridization signal is particularly noticeable when oligonucleotides or probes less than 100 nucleotides in length are used . 7. In the presence of 10 % dextran sulfate or 10 % polyethylene glycol , the rate of hybridization is ...
Contents
E | 8-8 |
SYNTHESIS OF THE FIRST STRAND OF CDNA 8 | 8-11 |
CONCENTRATING NUCLEIC ACIDS BY EXTRACTION WITH BUTANOL E | 8-16 |
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acrylamide agarose gel aliquots amplification annealing antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g cleavage cleaved cleotide coli DNA polymerase concentration containing ddNTPs deletions denatured digestion diluted dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers Maxam-Gilbert method MgCl2 microfuge tube minutes at 4°C molecules mRNA Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid piperidine plaques plasmid plasmid DNA plate polymerase chain reaction precipitation prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing gel sequencing reactions sonicated specific activity stored strand of cDNA synthesis T4 DNA polymerase target DNA target sequence template DNA termini transcription transfer Tris Cl pH vectors µg/ml