Molecular biology techniques: an intensive laboratory course
Saved in:
| Main Authors: | , |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
San Diego [u.a.]
Academic Press
1999
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | X, 234 S. . graph. Darst. |
| ISBN: | 0125839901 |
Staff View
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| 245 | 1 | 0 | |a Molecular biology techniques |b an intensive laboratory course |c Walt Ream and Katharine G. Field |
| 264 | 1 | |a San Diego [u.a.] |b Academic Press |c 1999 | |
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Record in the Search Index
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| adam_text | Contents
Preface ix
Course Synopsis 1
Introduction 2
Safety Precautions 7
Daily Schedule 9
Acknowledgments 15
EXERCISES:
I. DNA Preparation, Polymerase Chain
Reaction, and Molecular Cloning 17
A. Cesium chloride ethidium bromide density
gradient centrifugation 19
B. PCR to synthesize virD2 flanked with restriction
sites 27
C. Restriction digests of plasmid pGEX2 and
PCR products 35
D. Purification of DNA fragments from agarose 39
E. Ligation of PCR product to pGEX2 vector 45
F. Transformation of E. coli with the ligated
plasmid 49
G. Small scale preparation of plasmid DNA by the
alkaline lysis method 55
H. Restriction analysis 59
Study questions 63
v
CONTENTS
vi . . II. Protein Expression/ Purification/ and
Analysis 71
A. Expression and purification of a fusion protein 75
B. SDS polyacrylamide gel electrophoresis 81
C. Silver stain detection of proteins 87
D. Western blot (immunoblot) detection of proteins 91
Study questions 97
III. Oligonucleotide Directed Mutagenesis 99
A. Restriction digests of virD2 (in pCS64) and
PUC119 109
B. Purification of DNA fragments from agarose 111
C. Ligation of restriction fragment and vector 113
D. Transformation of E. coli with the ligated
plasmid and recovery of clones 117
E. Small scale preparation of plasmid DNA from
broth cultures 117
F. Restriction digest of DNAs: Examination to
confirm insert 117
G. Preparation of single stranded DNA template 119
H. Phosphorylation of oligonucleotide 125
I. Annealing mutant oligonucleotide to template 129
J. In vitro DNA synthesis by primer extension 133
K. Transform synthesis reaction into E. coli DH5a 137
L. Small scale preparation of plasmid DNA 137
M Confirmation of mutants by restriction analysis 138
Study questions 141
IV. DNA Sequencing 143
A. Polyacrylamide sequencing gel electrophoresis 147
B. Dideoxy sequencing 151
C. Automated sequencing 155
CONTENTS Vjj
D. Introduction to databases and gene sequence
analysis 157
Study questions 161
V. Southern Blot Detection of DNA 163
A. Preparation of genomic DNA from
Agrobacterium tumefaciens 165
B. Restriction digestion of genomic DNA 169
C. Agarose gel electrophoresis of restriction
fragments 171
D. Southern blot: Denaturation and blotting of DNA 175
E. Preparation of probe by nick translation 179
F. Hybridization and washing of Southern blots 185
Study questions 189
VI. Northern Blot Detection of mRNA 191
A. Preparation of RNA from tobacco leaves 195
B. Agarose formaldehyde gel electrophoresis 199
C. Northern blot: Denaturation and blotting of RNA 203
D. Probe preparation 207
E. Hybridization and washing of Northern blots 211
Study questions 215
VII. Protein Interaction Analysis in Yeast 217
A. Yeast transformation 221
B. Filter p galactosidase assay 225
Index 229
|
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| id | DE-604.BV012232809 |
| illustrated | Not Illustrated |
| indexdate | 2024-07-09T18:23:59Z |
| institution | BVB |
| isbn | 0125839901 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-008288876 |
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| physical | X, 234 S. . graph. Darst. |
| publishDate | 1999 |
| publishDateSearch | 1999 |
| publishDateSort | 1999 |
| publisher | Academic Press |
| record_format | marc |
| spelling | Ream, Walt Verfasser aut Molecular biology techniques an intensive laboratory course Walt Ream and Katharine G. Field San Diego [u.a.] Academic Press 1999 X, 234 S. . graph. Darst. txt rdacontent n rdamedia nc rdacarrier Biologie moléculaire - Manuels de laboratoire Moleküler biyoloji - Laboratuvar el kitapları Molecular biology Technique Methode (DE-588)4038971-6 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 s Methode (DE-588)4038971-6 s DE-604 Field, Katharine G. Verfasser aut HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008288876&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Ream, Walt Field, Katharine G. Molecular biology techniques an intensive laboratory course Biologie moléculaire - Manuels de laboratoire Moleküler biyoloji - Laboratuvar el kitapları Molecular biology Technique Methode (DE-588)4038971-6 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| subject_GND | (DE-588)4038971-6 (DE-588)4039983-7 |
| title | Molecular biology techniques an intensive laboratory course |
| title_auth | Molecular biology techniques an intensive laboratory course |
| title_exact_search | Molecular biology techniques an intensive laboratory course |
| title_full | Molecular biology techniques an intensive laboratory course Walt Ream and Katharine G. Field |
| title_fullStr | Molecular biology techniques an intensive laboratory course Walt Ream and Katharine G. Field |
| title_full_unstemmed | Molecular biology techniques an intensive laboratory course Walt Ream and Katharine G. Field |
| title_short | Molecular biology techniques |
| title_sort | molecular biology techniques an intensive laboratory course |
| title_sub | an intensive laboratory course |
| topic | Biologie moléculaire - Manuels de laboratoire Moleküler biyoloji - Laboratuvar el kitapları Molecular biology Technique Methode (DE-588)4038971-6 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| topic_facet | Biologie moléculaire - Manuels de laboratoire Moleküler biyoloji - Laboratuvar el kitapları Molecular biology Technique Methode Molekularbiologie |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008288876&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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