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Section: Genetics » Genetic Engineering and Biotechnology » Recombinant DNA and PCR
 
 
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  Genetic Engineering and Biotechnology 1. Recombinant DNA and PCR (Cloning and Amplification of DNA)
 
     
 
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Genetic Engineering and Biotechnology 1.  Recombinant DNA and PCR (Cloning and Amplification of DNA)
Restriction enzymes in cloning
Techniques used in recombinant DNA 
Cloning vectors for recombinant DNA
Plasmids as vectors
Bacteriophages as vectors
Plant and animal viruses as vectors
Transposons as vectors
Artificial chromosome vectors for cloning large DNA segments
Construction of chimeric DNA
Palindromes and staggered cleavage
Adding poly dA at the 3' ends of the vector and poly dT at the 3' ends of DNA clone
Blunt end ligation by T4 DNA ligase
Cloning in bacteria and eukaryotes
Cloning in bacteria
Cloning in eukaryotes
Molecular probes 
Labelling of probes
Applications of molecular probes
Construction and screening of genomic and cDNA libraries
Gene amplification : PCR and its applications
cDNA library from mRNA
Colony (or plaque) hybridization for screening of libraries
Gene Amplification : PCR and Its Applications
The basic polymerase chain reaction (PCR)
Different schemes of PCR
In recent years, techniques for manipulating prokaryotic as well as eukaryotic DNA have witnessed a remarkable development. This has allowed breakage of a DNA molecule at two desired places to isolate a specific DNA segment and then insert it in another DNA molecule at a desired position. The product thus obtained is called recombinant DNA and the technique often called genetic engineering. Using, this technique we can isolate and clone single copy of a gene or a DNA segment into an indefinite number of copies, all identical. This became possible because, bacteria, phages and plasmids reproduce in their usual manner even after insertion of foreign DNA, so that the inserted DNA will also replicate faithfully with the parent DNA. This technique is called 'gene cloning'. With this technique, genes can be isolated, cloned and characterized, so that the technique has led to significant progress in all areas of molecular biology. More recently, polymerase chain reaction (PCR) involving a thermostable DNA polymerase (Taq polymerase) has been used to obtain millions of copies of a DNA segment of choice. This PCR technique may eventually replace gene cloning in certain areas of research in the field of genetic engineering. In this section, we will discuss some details of these techniques involving production of recombinant DNA, and the cloning and amplification of DNA segments. The use of these techniques will be discussed in the next few topics of genetics on ePlantScience.com.

 
     






     
     
 
 
     
 
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