Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 57
Dilute an aliquot of the neutralized dNTP appropriately , and read the optical
density at the wavelengths given in the table ... Store in small aliquots at - 20°C .
Use strains C600 ( BNN93 ) for growth and BNN102 ( C600hfa ) for screening of
...
Dilute an aliquot of the neutralized dNTP appropriately , and read the optical
density at the wavelengths given in the table ... Store in small aliquots at - 20°C .
Use strains C600 ( BNN93 ) for growth and BNN102 ( C600hfa ) for screening of
...
Page 66
Remove two aliquots ( 2 ul 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 evaporation of the sample during subsequent
incubation .
Remove two aliquots ( 2 ul 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 evaporation of the sample during subsequent
incubation .
Page 65
In Falcon 2059 tubes , precooled to 0°C , combine 1 - and 5 - ul aliquots of the
undiluted reaction mixture and of each dilution of the reaction mixture with 200 -
ul aliquots of competent TG1 cells ( prepared as described in Chapter 1 , pages 1
.
In Falcon 2059 tubes , precooled to 0°C , combine 1 - and 5 - ul aliquots of the
undiluted reaction mixture and of each dilution of the reaction mixture with 200 -
ul aliquots of competent TG1 cells ( prepared as described in Chapter 1 , pages 1
.
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Contents
TEST LIGATION 3 29 | 3 |
SYNTHESIS OF THE FIRST STRAND OF cDNA 8 | 11 |
FRAGMENT OF E coli DNA POLYMERASE I 13 59 | 14 |
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Common terms and phrases
acid activity agarose aliquots allow amount amplification appropriate approximately bacteriophage bacteriophage T4 base buffer carried cDNA cells centrifugation Chapter chromatography cleaved clones colonies concentration constructed containing deletions denatured described determine digestion DNA polymerase dNTPs double-stranded efficiency electrophoresis et al ethanol expression Figure filters fragment gene genomic hybridization increase Incubate inserted interest isolated labeled length libraries ligation linear linkers method microfuge minutes mixture molecules mRNA mutagenesis mutagenic mutations Nucleic nucleotides obtained oligonucleotide original plaques plasmid plasmid DNA plate pool position possible prepared presence primer probe problem protein purified radiolabeled reaction recombinant region remove restriction enzyme room temperature samples screening sequence single single-stranded solution specific step Store strand synthesis target DNA target sequence template termini tion transfer transformants tube units usually vector volume