dec 6 dna libraries - A p p r o x im a te M a x im u m L e...

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Unformatted text preview: A p p r o x im a te M a x im u m L e n g th o f D N A T h a t C a n B e C lo n e d in V e c to r s Making DNA Libraries Vector Type Cloned DNA (kb) 20 25 45 100 300 1 000 S iz e o f fr a g m e n ts S iz e o f g e n o m e N u m b e r o f fr a g m e n ts n e e d e d D N A V S . R N A ( c D N A ) lib ra ry S c r e e n in g th e lib r a r y Plasmid Phage Cosmid P1 phage BAC (bacterial artificial chromosome) YAC (yeast artificial chromosome) C lo n in g H u m a n In s u lin intron intron 4 kb 333 bases 9 .5 k D a 1 17.9 F ig u r e 1 7 .9 C lo n in g H u m a n In s u lin intron intron 4 kb 333 bases 9 .5 k D a Size of human genome ~3.2 X 109 bp 3.2 X 106 kbp A v e r a g e s iz e o f in s e r t ~ 1 0 k b H o w m a n y c lo n e s m u s t b e s c r e e n e d P = 1-(1-F)N or N = ln(1-P)/ln(1-F) N = ln (1-0.99)/ ln (1-10/3.2X106) = ~3.3 x 106 clones Use of lac promoter for cloning * in s u lin g e n e 2 F ig . 9 .8 Insulin gene Lac PO a c DNA C lo n in g v e c to r d e s ig n e d fo r e x p r e s s io n o f th e p r o te in T a g g e d a n tib o d y s p e c ifi c fo r th e p r o te in p r o d u c e d B o u n d a n tib o d y d e te c te d b y X -r a y fi lm (if r a d io a c tiv e ), e n z y m e a s s a y , fl u o r e s c e n c e o r o th e r ta g d e te c tio n 3 ...
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This note was uploaded on 02/22/2010 for the course BIO 51980 taught by Professor Buskirk during the Fall '06 term at University of Texas at Austin.

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