Gene Cloning.transformation

Gene Cloning.transformation - Cleavage produces cohesive...

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Plasmid/Gene Cloning (ignore the agar part. . I didn’t get it too much) DNA cloning is a method of isolating a particular sequence of DNA from a complex mixture of  DNA sequences. You clone by inserting DNA in a plasmid vector, usually a phage, because it can insert the  genetic material in a cell and replicate.  Plasmid Vector must have 3 elements: Cloning Site- where restriction enzyme cuts and foreign DNA is inserted. Drug Resistant gene- which will destroy antibodies to allow selective growth Replication Origin- to allow plasmid to replicate in host cell 1 st Restriction enzymes  cleaves Plasmids and also cleaves the desired DNA. (Because its cleaved by the same  restriction enzyme, they will have the same  conformation,  and DNA will be able to be inserted/hybridize into the plasmid)
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Unformatted text preview: Cleavage produces cohesive ends/staggered ends/ “sticky ends” 2 nd- Sticky ends officially hybridize with the foreign DNA that was fragmented by the same restriction enzyme. The hybridizing gets sealed by DNA Ligase- forming phosphodiester bonds 3 rd- by Transformation , plasmids are incorporated in bacterial host cells Each cell has different plasmids b/c each plasmid has different/unique DNA That is because during Transformation, the repair system makes sure that there are no nucleotide sequence differences of the donor/recipient DNA; therefore some cells get repaired and keep the ‘ampicillin resistance’, while others get rid of the ‘amipicillin resistance’. Transformation Refresh ….lol
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This note was uploaded on 04/18/2008 for the course MICRO BIO 132 taught by Professor Ray during the Spring '08 term at Long Island U..

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Gene Cloning.transformation - Cleavage produces cohesive...

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