Molecular cloning: a laboratory manual, Volume 2 |
From inside the book
Results 1-5 of 14
Page 8-29
... ligation ) . A single centrifugation step through a Centricon - 100 membrane removes ~ 95 % of the primers and deoxynucleotides from the PCR . This process also inactivates the thermostable DNA poly- merase ( presumably by absorption to ...
... ligation ) . A single centrifugation step through a Centricon - 100 membrane removes ~ 95 % of the primers and deoxynucleotides from the PCR . This process also inactivates the thermostable DNA poly- merase ( presumably by absorption to ...
Page 8-30
... ligase , the cloning efficiency of blunt- ended DNA is 10-100 - fold lower than the efficiencies attained with DNAS equipped with cohesive termini . In addition , blunt - end ligation provides no opportunity to direct the orien- tation ...
... ligase , the cloning efficiency of blunt- ended DNA is 10-100 - fold lower than the efficiencies attained with DNAS equipped with cohesive termini . In addition , blunt - end ligation provides no opportunity to direct the orien- tation ...
Page 8-31
... ligation of cohesive ends . • Using a linearized plasmid vector fitted with a protruding 3 ' thymidylate residue at each of its 3 ' termini . These T vectors are used to clone DNA fragments that carry a nontemplated , unpaired ...
... ligation of cohesive ends . • Using a linearized plasmid vector fitted with a protruding 3 ' thymidylate residue at each of its 3 ' termini . These T vectors are used to clone DNA fragments that carry a nontemplated , unpaired ...
Page 8-32
... ligation of the plasmid to a target DNA molecule destroy the restriction site . This prevents the restriction enzyme from destroying recombinants generated during the ligation reaction . The net effect of constant reclamation of unit ...
... ligation of the plasmid to a target DNA molecule destroy the restriction site . This prevents the restriction enzyme from destroying recombinants generated during the ligation reaction . The net effect of constant reclamation of unit ...
Page 8-33
... ligation of the target fragment to the vector should not regenerate the restriction site . Among the plasmid vectors that are commonly used for cloning of blunt - ended PCR products are the Bluescript - type plasmids and plasmids ...
... ligation of the target fragment to the vector should not regenerate the restriction site . Among the plasmid vectors that are commonly used for cloning of blunt - ended PCR products are the Bluescript - type plasmids and plasmids ...
Contents
8-6 | |
8-17 | |
8-23 | |
Analysis of Interacting Proteins with Surface Plasmon Resonance Spectroscopy 18 | 8-96 |
INFORMATION PANELS | 8-107 |
INFORMATION PANELS | 8-109 |
Chapter 9 | 8-120 |
Preparation of Radiolabeled DNA and RNA Probes 9 1 | 8-128 |
Exon Trapping and Amplification 11 80 | 11-80 |
INFORMATION PANELS | 11-98 |
Chain Reaction | 11-125 |
Chapter 1 | 12-1 |
Chapter 4 | 12-4 |
Chapter 12 | 12-12 |
Chapter 16 | 12-16 |
INTRODUCTION | 12-32 |
Chapter 15 | 9-15 |
Chapter 18 | 9-18 |
INTRODUCTION | 9-25 |
PROTOCOLS | 9-33 |
INFORMATION PANELS | 9-76 |
1 | 10-1 |
Chapter 10 | 10-10 |
Volume 1 | 10-11 |
Volume 2 | 10-16 |
INFORMATION PANELS | 10-43 |
Chapter 3 | 11-3 |
Chapter 5 | 11-5 |
Chapter 11 | 11-7 |
Chapter 8 | 11-8 |
PROTOCOLS | 11-13 |
INFORMATION PANELS | 12-94 |
1 | 12-115 |
Chapter 13 | 13-13 |
INFORMATION PANELS | 13-63 |
Purification of Closed Circular DNA by Equilibrium Centrifugation in CsClEthidium 1 69 | 13-69 |
INFORMATION PANELS | 14-1 |
Chapter 2 | 14-2 |
Chapter 14 | 14-3 |
INFORMATION PANELS | 14-9 |
INFORMATION PANELS | 7 |
1 | 1-1 |
Chapter 7 | 1-7 |
Eukaryotic Cells | 1-17 |
Other editions - View all
Molecular Cloning: Selected Applications in Medicine and Biology Gregory G. Brown Limited preview - 2011 |
Common terms and phrases
Acad agarose gel aliquots amplified DNA annealing Appendix bacteriophage M13 bands base Biol BioTechniques catalyzed cDNA clones cDNA libraries cells centrifuge Chapter cleavage concentration containing cycle ddNTP denatured detection digestion DNA fragments DNA sequencing DNA templates dNTPs double-stranded DNA efficiency Escherichia coli ethanol exonuclease gel electrophoresis gene H₂O hybridization information panel Klenow fragment labeled ligation method microfuge tube microtiter plate minutes molecules mRNA mutations Natl Nucleic Acids Nucleic Acids Res nucleotides oligonucleotide oligonucleotide primers PCR products phagemid plaques plasmid plasmid DNA pmoles polyacrylamide gel polymerase chain reaction polymerization polynucleotide kinase preparation probes protein Protocol purified radioactivity radiolabeled reaction mixture reagents listed recombinant requires the reagents residues restriction enzyme reverse transcriptase RNase room temperature samples screening Sequenase sequencing gel sequencing reactions single-stranded DNA Step strand synthesis target DNA template DNA termini thermal cycler thermostable DNA polymerase tion transcription vector vitro wild-type µg/ml
Popular passages
Page 9-91 - Melton, DA, Krieg, PA, Rebagliati, MR, Maniatis, T., Zinn, K., and Green, MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 12, 7035-7056.
Page 14-37 - Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
Page 9-88 - Eldeiry, WS, Tokino, T., Velculescu. VE, Levy, DB, Parsons. R., Trent, JM, Lin, D., Mercer, WE, Kinzler, KW, and Vogelstein, B.
Page 9-92 - Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase.
Page 9-87 - Campbell, AP, Chenchik, A., Moqadam, F., Huang, B., Lukyanov, S., Lukyanov, K., Gurskaya, N., Sverdlov, ED, and Siebert, PD (1996).
Page 8-120 - Gilliland. G., Perrin. S., Blanchard. K., and Bunn, HF (1990) Analysis of cytokine mRNA and DNA detection and quantitation by competitive polymerase chain reaction.
Page 8-120 - Frohman, MA, Dush, MK, and Martin, GR (1988) Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc. Natl. Acad. Sci. USA 85, 8998-9002. 2. Edwards, JBDM, Delort, J., and Mallet, J. (1991) Oligodeoxyribonucleotide ligation to single-stranded cDNAs: A new tool for cloning 5' ends of mRNAs and for constructing cDNA libraries by in vitro amplification.
Page 9-87 - DV, Angerer, LM, and Angerer, RC (1984) Detection of mRNAs in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev. Biol. 101, 485-502.
Page 12-103 - Birnboim, HC and Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
Page 9-88 - A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.