Cloning in Phage Lamda Vectors, Subcloning DNA Fragments, Propagated Vectors, Construction Genomic, Cloning Phage, Lamda Restriction Enzyme

Scholarships Dictionary Visa Details Study Abroad Exam Results Online Test Jobs

	Home
Chimeric DNA Molecular Probes and Gene Libraries Chimeric DNA Molecular Probes and Gene Libraries Introduction Restriction Enzymes for Cloning Technique of Restriction Mapping Restriction Clevage and Gel Electrophoresis Construction of a Restriction Map Use of Partial Digests end Labelling and Hybridization in Restriction Mapping Construction of Chimeric DNA Cloning of Plasmid Vectors Directional Cloning of DNA Fragments with Heterologus Protruding Termini Adding Poly dA at 3' Ends of DNA Clone Blunt End Ligation by T4 DNA Ligase Cloning in Phage λ and Cosmid Vectors Cloning in phage λ Vectors Cloning in Cosmid Vectors Hosts for Cloning Vectors Bacteria as Hosts Eukaryotes as Hosts Molecular Probes Preparation of Probes Genomic DNA Probes

Home >> Biotechnology and Genomics >> Chimeric DNA Molecular Probes and Gene Libraries >> Cloning in Phage λ Vectors

Your Ad Here

Cloning in phage λ and cosmid vectors

Cloning in phage λ. vectors. Phage λ vectors are used for a variety of purposes including the following: (i) subcloning of DNA fragments that are propagated in vectors with large capacity (e.g. cosmids) and (ii) construction of genomic and cDNA libraries. Cloning in phage λ vectors involves the following steps: (i) digestion of phage λ with a restriction enzyme (digestion with two enzymes, one followed by the other is sometimes carried out to remove the stuffer;

this is necessary in case of only some of the replacement vectors) and purification of phage λ; purified arms of several popular A vectors including those for EMBL3 and EMBL4, and those for λgt10 and λgt11 are also available commercially, so that if the arms are procured commercially, this step involving digestion and purification can be dispensed with; (ii) ligation of A arms to fragments of foreign DNA to be cloned;

(iii) packaging of recombinant phage A in vitro, utilizing the packaging -extract that is available commercially or can be prepared in the laboratory (packaging extract is a lysate of phage A infected E. coli cells and contains empty phage heads, unattached phage tails and phage-encoded proteins needed for DNA packing); this is followed by infection of E.coli competent cells by packaged recombinant

A vectors, although rarely transfection with naked A DNA without packaging can also be tried; (iv) multiplication of transformed E. coli cells and (v) lysis of E. coli cells to release recombinant phage, forming plaques and recovery of cloned DNA segment.

	Home
c DNA Probes Synthetic Oligonucleotides as Probes RNA Probes or Riboprobes Labelling of Probes Radiolablled Probes Non Radioactive Probes Biotin Labelled Probes Digoxigenin Labelled Probes Alternatives to Biotin and Digoxigenin Labelling Amplification of DNA Probe Signals Techniques Used in Molecular Probing Southern Nothern and Western Blotting Southern Blotting Northern Blotting Western Blotting Dot Blots and Slot Blots Applications of Molecular Probes Construction and Screening of Genomic and cDNA Libraries Genomic Library by Shotgun Experiment cDNA Library from mRNAs Colony or Plaque Hybridization for Screening of Libraries Chromosome Walking and Characterization of Chromosome Segments Chromosome Jumping or Hopping Libraries BAC Libraries and Assembly of BACs into Contigs