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Academic Report

Zhongdi Song

Date:2012-10-20

Contents

Gene Konckout

Colony PCR

Phenotypic Screening

Gene konckout

Eight genes sll1734, sll1549, sll0258, sll0057

slr1339, slr1472, ssr1853,ssr3402

Basic operation

primer design

PCR(1kb)-electrophoresis-purification-fusion PCR(3kb)--

transformation-tranfer-colony PCR-enrichment--

culture in the liquid medium-conservation or further study

PCR(1Kb)

1,2—sll1734; 3,4—slr1339; 5,6—slr1472; 7,8—sll0258

PCR(1Kb)

1,2—ssr1853; 3,4—ssr3402; 5,6—sll1549; 7,8—sll0057

Fusion PCR(3Kb)

1—sll1734 ； 2—slr1339 ； 3—slr1472 ； 4—sll0258

5—ssr1853 ； 6—ssr3402 ； 7—sll1549 ； 8—sll0057

Colony PCR

Colony PCR

1—3056 ， 3394(338bp) ； 2—2952,2487(465bp) ； 3—501bp ； 5—3208,2965(243bp) ； 6—2996,2327(669bp) ； 7—3138,2928(210bp) ； 8—2959,2482(477bp)

Colony PCR

sll1549—3208,2965(243bp) ； sll0057—3138,2928(210bp) sll0258—2959,2482(477bp) ； ssr1853—3073,2401(672bp)

Colony PCR(10.6)

Colony PCR

Influencing factors

PCR ：① the concentration of template

②cycle numbers

electrophoresis ： ① electrophoresis buffer

②voltage

③gel

④sample volume

Phenotypic screening

96-well plates

33 mutants

16 stress conditions

eg. 3HP-0.46mM

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