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- shortened
- lengthened
- copied into new DNA molecules
- modified by the addition or removal of specific chemical groups
♦ Restriction endonucleases
♦ Ligases
Nucleases- degrade DNA molecules by breaking the
phosphodiester bonds that link one nucleotide to the next in a
DNA strand
X
Klenow fragment
Taq polymerase from Thermus aquaticus is used in the
polymerase chain reaction (PCR). This enzyme is thermostable,
meaning that is resistant to denaturation by heat treatment
Other polymerases proofreading (3´-5´exonuclease) used:
Host-controlled restriction
and modification of phage
l in E. coli strain K, by
efficiency of plating (EOP)
Enzymes for cutting DNA- Restriction endonucleases
♦ Ligation is performed at 16 ºC
♦ Ligation of complementary sticky ends is much more
efficient, because the base-paring between the two
molecules is a relatively stable structure for the enzyme
to work on
Putting sticky ends onto a blunt-ended molecule
Adaptors-
short synthetic
oligonucleotides with
one sticky end
Producing sticky ends by homopolymer tailing
(Cont.)