Molecular cloning

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When the hybridization is complete, remove the hybridization solution and immediately immerse the filters in a large volume (300-500 ml) of Wash solution 1 at room temperature. Agitate the filters gently and turn them over at least once during washing. After 5 minutes, transfer the filters to a fresh batch of wash solution and continue to agitate them gently. Repeat the washing procedure twice more. 5 hours in 300-500 ml of Wash solution 2 at 68°C. Dry the filters in the air at room temperature on 3MM paper.

1987. Transformation of Lactobacillus casei by electroporation. FEMS Microbiol. Lett. 44:173-177. 2. , and Flickinger J. 1988. Transformation of bacteria by electroporation. Trends Biotechnol. 6:303-309. 3. W. 1988. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 16:6127-6145. pronumber=26&chpnumber=1 (1 / 2) [2002-2-18 16:15:27] Chapter:1 Protocol:27 Screening Bacterial Colonies Using X-gal and IPTG: ±-Complementation CHAPTER 1 > PROTOCOL 27 printer friendly version Protocol 27 Screening Bacterial Colonies Using X-gal and IPTG: -Complementation -complementation occurs when two inactive fragments of E.

LB, YT, or Terrific Broth) containing the appropriate antibiotic. After a period of growth, plasmid DNA can be isolated from the culture by one of the minipreparation methods described in Chapter 1, Protocol 1 and Chapter 1, Protocol 4 and can be further analyzed by restriction endonuclease digestion or by PCR. REFERENCES 1. Grunstein M. S. 1975. Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. 72:3961-3965. pronumber=32&chpnumber=1 (1 / 2) [2002-2-18 16:17:44] Cold Spring Harbor Laboratory Press - Molecular Cloning - Chapter 2 Chapter 2 Bacteriophage and Its Vectors Protocol 1: Plating Bacteriophage This protocol describes a method for generating isolated plaques from a stock of bacteriophage .

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