Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-66
... EcoRI methylase is inhibited by Mg ++ ions . 2. Remove two aliquots ( 2 μl each ) and place in small ( 0.5 - ml ) microfuge tubes . Number these aliquots 1 and 2. Store at 0 ° C . It is important to use small microfuge tubes to minimize ...
... EcoRI methylase is inhibited by Mg ++ ions . 2. Remove two aliquots ( 2 μl each ) and place in small ( 0.5 - ml ) microfuge tubes . Number these aliquots 1 and 2. Store at 0 ° C . It is important to use small microfuge tubes to minimize ...
Page 8-80
... EcoRI sites within the cDNA inserts is less than 1 site per 4 kb . This problem arises when methylation of EcoRI sites within the double- stranded cDNA is not complete . In this case , it is usually necessary to check the methylation ...
... EcoRI sites within the cDNA inserts is less than 1 site per 4 kb . This problem arises when methylation of EcoRI sites within the double- stranded cDNA is not complete . In this case , it is usually necessary to check the methylation ...
Page 15-32
... EcoRI 5 ' - 85 BamHI restriction and recombination with DNA ligase ACAAACCCCGCCCAGCCGGATCCGGTTGGCGAATTCGAACACGCAGATGC BamHI linker EcoRI ACAAACCCCGCCCAGCGTCTTGTCATTGGCGAATTCGAACACGCAGATGC EcoRI LS -95 / -85 Wild type FIGURE 15.5 ...
... EcoRI 5 ' - 85 BamHI restriction and recombination with DNA ligase ACAAACCCCGCCCAGCCGGATCCGGTTGGCGAATTCGAACACGCAGATGC BamHI linker EcoRI ACAAACCCCGCCCAGCGTCTTGTCATTGGCGAATTCGAACACGCAGATGC EcoRI LS -95 / -85 Wild type FIGURE 15.5 ...
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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 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 µg/ml