Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-15
... strand was a double - stranded cDNA molecule closed at the terminus corresponding to the 5 ' terminus of the mRNA by a hairpin loop . This loop was then digested with the single - strand- specific nuclease S1 to give a double - stranded ...
... strand was a double - stranded cDNA molecule closed at the terminus corresponding to the 5 ' terminus of the mRNA by a hairpin loop . This loop was then digested with the single - strand- specific nuclease S1 to give a double - stranded ...
Page 8-64
... Strand of cDNA [ The unlabeled first - strand reaction needs no further treatment before pro- ceeding to the synthesis of the second strand of cDNA . [ 1. Add the following reagents directly to the first - strand reaction mixture : 10 ...
... Strand of cDNA [ The unlabeled first - strand reaction needs no further treatment before pro- ceeding to the synthesis of the second strand of cDNA . [ 1. Add the following reagents directly to the first - strand reaction mixture : 10 ...
Page 8-65
... strand of cDNA and to one containing the second strand of cDNA . Incubate all four tubes for 30 minutes at 37 ° C . g . Analyze the size of first- and second - strand products by electrophoresis through an alkaline 1 % agarose gel ( see ...
... strand of cDNA and to one containing the second strand of cDNA . Incubate all four tubes for 30 minutes at 37 ° C . g . Analyze the size of first- and second - strand products by electrophoresis through an alkaline 1 % agarose gel ( see ...
Contents
E | 8-8 |
SYNTHESIS OF THE FIRST STRAND OF CDNA 8 | 8-11 |
CONCENTRATING NUCLEIC ACIDS BY EXTRACTION WITH BUTANOL E | 8-16 |
Copyright | |
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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