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Survey of Molecular Techniques

Survey of Molecular Techniques

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Published by Dozdi

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Published by: Dozdi on Apr 14, 2010
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11/02/2012

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Survey of molecular techniques
Enzymes for DNA manipulation
DNA polymerasesnucleasesligasesend-modification enzymes
 
DNA polymerases(template-dependent)
synthesize new DNA chains (complementary to template)require oligonucleotideprimersmay possess 3’-5’ exonuclease activity (for proofreading)some forms are thermostable (e.g.,
Taq
polymerase)some forms require RNA as template (RNA polymerases),synthesizing cDNAs
Nucleases
hydrolyze phosphodiester bonds in nucleic acidssome are specific for DNA, others for RNAmain types:exonucleaseendonuclease
 
Exonucleases: examples
ExonucleaseIII -removes residues from 3’ endsof a DNA strandBacteriophage
λ
exonuclease -removes residuesfrom 5’ ends of a DNA strand
Endonucleases
cut at internal phosphodiesterbondsrestriction enzymes -an important class of DNAendonucleases; play a central role inrecombinant DNA technology

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