Part II Strategies for Gene Isolation and Characterization
Chapter 16
Cloning Strategies: An Overvierw
FRIEDHELM HILDEBRANDT ••••••••••.....•.•.•.••.••.••••• 257
Chapter 17
Genomic Libraries
LUITZ F. ONUCHIC AND GREGORY G. GERMINO .•.......••.•••• 261
Chapter 18
cDNA Libraries
LUITZ F. ONUCHIC AND GREGORY G. GERMINO ••.•.••..•.••••. 273
Chapter 19
cDNA Library Screening
JOHANN PETER HOSSLE .•.•••..•••..•.•.••................ 288
Subprotocol 1: Platingof a A.gt11 cDNA Library ................ 296
Subprotocol2: Hybridization Screening- cDNA Sequences as Probes 298
Subprotocol 3: Immuno-Screening ........................... 300
Subprotocol 4: Purification of Phage Particles, DNA Isolation
and Subcloning ............................. 306
Chapter 20
Expression Cloning with Xenopus Oocytes
DANIEL MARKOVICH, ANDREAS WERNER AND HEINI MURER •.•..• 310
Chapter 21
Transgenie Animals
ECKART SCHOTT, MARTIN PAUL AND DETLEV GANTEN . . . . . . . . . . 319
Chapter 22
Expression in Eukaryotic Cells: Gene Transfer Using Retroviral Vectors
MARTIN MARX ..•.••....•••••••••..••.••..•..•..••••.•.• 337
Subprotocol 1: Synthesis of the Insert ........................ 339
Subprotocol 2: Purification ................................ 340
Subprotocol 3: Ligation of the Insert into the V ector ............ 340
Subprotocol 4: Transfeetion of E. coli HB101 and Plasmid
DNA Isolation .............................. 341
Subprotocol 5: Assay for the Presence of the DNA Insert ......... 343
Contents
XI