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Academic Report

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  1. Academic Report Zhongdi Song Date:2012-10-20

  2. Contents Gene Konckout Colony PCR Phenotypic Screening

  3. 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

  4. PCR(1Kb) 1,2—sll1734; 3,4—slr1339; 5,6—slr1472; 7,8—sll0258

  5. PCR(1Kb) 1,2—ssr1853; 3,4—ssr3402; 5,6—sll1549; 7,8—sll0057

  6. Fusion PCR(3Kb) 1—sll1734;2—slr1339;3—slr1472;4—sll0258 5—ssr1853;6—ssr3402;7—sll1549;8—sll0057 Colony PCR

  7. 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)

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

  9. Colony PCR(10.6)

  10. Colony PCR • Influencing factors PCR:①the concentration of template ②cycle numbers electrophoresis: ①electrophoresis buffer ②voltage ③gel ④sample volume

  11. Phenotypic screening • 96-well plates • 33 mutants • 16 stress conditions

  12. eg. 3HP-0.46mM

  13. Thank You !