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Lab meeting 2014.02 .24

Lab meeting 2014.02 .24. Noh ji eun. Calreticulin. PCR purification - AccuPrep PCR purifiction kit -Components. Calreticulin. Big dye reaction <Big dye mixture>. Calreticulin. Big dye reaction <Big dye condition> 96 ℃ – 96 ℃ – 50 ℃ – 60 ℃ – 22 ℃

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Lab meeting 2014.02 .24

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  1. Lab meeting2014.02 .24 Noh jieun

  2. Calreticulin • PCR purification -AccuPrep PCR purifiction kit -Components

  3. Calreticulin • Big dye reaction <Big dye mixture>

  4. Calreticulin • Big dye reaction <Big dye condition> 96 ℃ – 96 ℃ – 50 ℃ – 60 ℃ – 22 ℃ 1min 10sec 5sec 4min ∞ └ 25cycle ┘

  5. Calreticulin • Big dye purification <Big dye purification mixture> 그 외 필요한 시약 -70%ethanol 350㎕ -Hi-Di-Formamide 15㎕

  6. Calreticulin • Sequencing Result -CALR1,2,3,5,6,7,8,9,10,11,12,14,15 :mutation 없음. -CALR4,13 :Sequencing Result 나오지않음.

  7. Calreticulin • PCR reaction

  8. Calreticulin • PCR Condition

  9. Calreticulin • PCR Result → Electrophoresis 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

  10. Calreticulin • Gene scan • Gene scan PCR condition : 95℃ – 4℃ 5min ∞

  11. Calreticulin • Gene scan result :CALR 1,2,3,6,8,9,10,11,12,14-mutation 없음 CALR 5,7-mutation 있음

  12. Calreticulin • Sample: MPN환자에서 2011년도 2명, 2012년도 45명 JAK2 (-) 환자.

  13. Calreticulin • PCR Result → Electrophoresis 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47

  14. Thank You

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