Screening a cDNA Library
来源：本站原创 2006/7/28 17:01:00
Screening a cDNA Libraryfor use with HybriZAP zebrafish cDNA libraries
ObjectivecDNA library screening allows detection of expressed genes for subsequent cloning and sequencing.
Selecting a LibraryCurrently, our lab has access to cDNA libraries from three developmental time points.
Choose the appropriate library for when you expect your message of interest is expressed.
- Maternal: messages from embryos before midblastula-transistion, when zygotic transcription begins (~0-3 hours of development).
- Shield: messages from the shield stage, which signifies the beginning of gastrulation (~6 hours of development).
- Post-somitegenesis: messages after the formation of somites is complete (~24 hours of development).
Overview of Procedures
- Titer the library to determine the concentration of infectious phage.
- Plate and lift the phage particles.
- Hybridize the immobilized cDNA with labeled probe.
- Pick positive plaques.
- Cycle through steps 2-4 until a single cDNA containing phage is isolated, generally three cycles.