Lambda gt11/EcoR I/CIAP-Treated Vector Kit Instruction Manual, this manual introduces the materials, storage conditions, introduction, preparation of host strains, host strain genotypes, growth and maintenance of host strains, preparation of -80°C bacterial glycerol storage solution.
This document provides instructions for using the Lambda gt11/EcoR I/CIAP-Treated Vector Kit (Catalog #234211) to construct cDNA libraries. It details the materials provided, including the digested vector, test insert, host strains (Y1088, Y1089r-, Y1090r-), and packaging extract.
The guide covers essential procedures such as preparing and maintaining host bacterial strains, performing color selection using IPTG-X-gal to identify recombinant plaques, ligating inserts into the vector, and packaging the library. It also includes step-by-step instructions for amplifying the library over three days, performing plaque lifts, hybridizing with probes or antibodies for screening, and troubleshooting common issues.
| Insert Size Capacity | up to 7.2 kilobases |
|---|---|
| Vector Quantity (Catalog #234211) | 10 pg |
| Test Insert Quantity (Catalog #234211) | 1.25 wg |
| Packaging Extract Volume | 11 x 25 ul |
The vector should be stored at -20°C after thawing and aliquoting.
The Y1088 host strain gives a high lambda phage yield and is ideal for this purpose.
Recombinant plaques appear white (clear) while background plaques are blue when plated with IPTG and X-gal.