Lambda ZAP® II Premade Library INSTRUCTION MANUAL
The document serves as an instruction manual for the Lambda ZAP II Premade Library, a premade cDNA library constructed in the Lambda ZAP II vector. It details the materials provided with the kit, including host strains (XL1-Blue MRF' and SOLR) and helper phages (ExAssist and VCSM13), along with specific storage conditions requiring -80°C.
The guide covers essential laboratory protocols for working with the library, including establishing bacterial stocks, determining background via color selection, titering helper phage and the library itself, performing plaque lifts, hybridizing and screening clones, and in vivo excision of the pBluescript SK(-) phagemid. It also includes recipes for preparing necessary media and reagents.
| Vector System | Lambda ZAP II |
|---|---|
| Host Strain 1 | XL1-Blue MRF' |
| Host Strain 2 | SOLR™ strain |
| Helper Phage 1 | ExAssist® interference-resistant helper phage |
| Helper Phage 2 | VCSM13 interference-resistant helper phage |
| Storage Temperature | -80°C |
The premade library should be stored at -80°C.
You should use the SOLR™ strain for plating excised phagemids.
Stratagene recommends VCSM13 interference-resistant helper phage for single-stranded rescue.
You need Isopropyl-1-thio-B-D-galactopyranoside (IPTG) and 5-Bromo-4-chloro-3-indoyl-B-D-galactopyranoside (X-gal).