LEC16 - 11/6/2008 Announcements Last day for regrades You...

Info iconThis preview shows pages 1–7. Sign up to view the full content.

View Full Document Right Arrow Icon
11/6/2008 1 Announcements Last day for regrades You may pick up regraded exams next week in the LS Core office Vectors Classic vectors pBR322 First artificial plasmid Named after scientist Bolivar and Rodriguez Derived from 2 naturally occurring plasmids pUC19 Introduced lacZ fragment and multicloning site
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
11/6/2008 2 Insertion of DNA between BamHI and SalI sites results in amp-r, tet-s plasmid pUC vectors introduced lacZ alpha fragment and multiple restriction sites
Background image of page 2
11/6/2008 3 Multi-cloning site (MCS) Insertion into any of these sites will disrupt lacZ gene: white colony Options for cloning genes PCR amplify gene directly from the genomic DNA and then insert this genomic DNA and then insert this fragment into a vector Need knowledge of sequence of gene of interest Screen or select a clone from a library of clones Order directly from facility
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
11/6/2008 4 Cloning a PCR product Gene X PCR product Digest EcoR I Ligate I linkers ORI I Digest I Mix, ligate Gene X amp r I I amp r ORI Use PCR to check orientation of insert Gene X I I ORI X eneG I I ORI amp r amp r No product
Background image of page 4
11/6/2008 5 Screen or select a clone from a library of clones Library of recombinant clones Collection of recombinant molecules representing all the genomic DNA or expressed genes in a cell or organism Since this is a mixture of recombinant molecules, one must screen or select for gene of interest
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
11/6/2008 6 Two types of libraries: Genomic library Pieces of the enome inserted into a vector
Background image of page 6
Image of page 7
This is the end of the preview. Sign up to access the rest of the document.

This note was uploaded on 07/08/2009 for the course LIFESCI LS 3 taught by Professor Paulolague during the Fall '09 term at UCLA.

Page1 / 19

LEC16 - 11/6/2008 Announcements Last day for regrades You...

This preview shows document pages 1 - 7. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online