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酵母遗传实验方法 冷泉港实验课手册 英文版PDF|Epub|txt|kindle电子书版本网盘下载
- (美)Dan Burke等著 著
- 出版社: 北京:清华大学出版社
- ISBN:7302050961
- 出版时间:2002
- 标注页数:205页
- 文件大小:7MB
- 文件页数:221页
- 主题词:
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图书目录
Experiments1
Ⅰ.Looking at Yeast Cells1
Ⅱ.Isolation and Characterization of Auxotrophic,Temperaturesensitive,and UV-sensitive Mutants7
Ⅲ.Meiotic Mapping17
Ⅳ.Mitotic Recombination and Random Spore Analysis29
Ⅴ.Transformation of Yeast39
Ⅵ.Synthetic Lethal Mutants49
Ⅶ.Gene Replacement55
Ⅷ.Isolation of ras2 Suppressors73
Ⅸ.Manipulating Cell Types79
Ⅹ.Isolating Mutants by Insertional Mutagenesis89
Ⅺ.Two-hybrid Protein Interaction Method97
Techniques and Protocols103
1.High-efficiency Transformation of Yeast103
2.“Lazy Bones”Plasmid Transformation of Yeast Colonies107
3.Yeast DNA Isolations109
A.Yeast DNA Miniprep(40 ml)109
B.Yeast DNA Miniprep(5 ml)110
C.A Ten-minute DNA Preparation from Yeast111
D.Yeast Genomic DNA:Glass Bead Preparation113
4.Yeast Protein Extracts115
5.Yeast RNA Isolation117
6.Hydroxylamine Mutagenesis of Plasmid DNA121
7.Assay of β-Galactosidase in Yeast123
8.Plate Assay for Carboxypeptidase Y127
9.Random Spore Analysis129
10.Yeast Vital Stains131
11.Yeast Immunofluorescence133
12.Actin Staining in Fixed Cells137
13.PCR Protocol for PCR-mediated Gene Disruption139
14.Yeast Colony PCR Protocol141
15.Measuring Yeast Cell Density by Spectrophotometry143
16.Cell Synchrony145
17.Chromatin Immunoprecipitation149
18.Flow Cytometry of Yeast DNA155
19.Logarithmic Growth157
20.EMS Mutagenesis159
21.Tetrad Dissection161
22.Making a Tetrad Dissection Needle165
23.Picking Zygotes167
24.Determining Plating Efficiency169
Appendices171
A.Media171
B.Stock Preservation183
C.Yeast Genetic and Physical Maps185
D.Templates for Making Streak Plates193
E.Electrophoretic Karyotypes of Strains for Southern Blot Mapping195
F.Strains197
G.Counting Yeast Cells with a Standard Hemocytometer Chamber199
H.Tetrad Scoring Sheet201
I.Trademarks203
J.Suppliers205