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[»ý¹°ÇÐ º¸°í¼] DNA Isolation, PCR, Àü±â¿µµ¿ ½ÇÇè |
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[»ý¹°ÇÐ º¸°í¼] DNA Isolation, PCR, Àü±â¿µµ¿ ½ÇÇè |
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<-Date><> <-Title><¨çDNA Isolation ¨èPCR(polymerase chain reaction) ¨éÀü±â¿µµ¿(electrophoresis)> <-Object ><¨ç¹ÚÅ׸®¾Æ·ÎºÎÅÍ DNA¸¦ ºÐ¸®ÇØ ³½´Ù. ¨èƯÁ¤ DNA ºÎÀ§¸¦ ¹Ýº¹ ÇÕ¼ºÇÏ¿© ¿øÇÏ´Â DNA ºÐÀÚ¸¦ ÁõÆø ½ÃŲ´Ù. ¨éÀü±â¿µµ¿Àº DNA¸¦ Å©±â¿¡ µû¶ó¼ ºÐ¸®ÇÏ´Â ¹æ¹ýÀÌ´Ù.> <-Materials& Methods ><¨çCell lysis sol, proteinase K, RNase, PPT sol, e-tube, DNA binding sol, spin colmn, column washing sol, water, ¿ø½ÉºÐ¸®±â 1. Cell lysis sol 300§¡ (inverting) 2. proteinase K 10§¡ (inverting) 56¡É¿¡¼ 15ºÐ 3. RNase A 3§¡ 37¡É¿¡¼ 15ºÐ 4. PPT sol 100§¡¡æ Vortexing¡æ In ice¿¡¼ 5ºÐ 5. 10ºÐ µ¿¾È 1300rpm centrifuge 6. »óµî¾× 390§¡¡æ e-tube¡æ DNA binding sol 600§¡ 7. 650§¡ in Spin column¡æ 1ºÐ µ¿¾Ècentrifuge 8. Column washing sol 600§¡¡æ 1ºÐ µ¿¾Ècentrifuge 9. Column washing sol 500§¡¡æ 1ºÐ µ¿¾Ècentrifuge 10. spin columnÀ» »õ e tube¿¡ ¿Å±â°í water 100§¡¸¦ ³Ö°í 5ºÐ Á¤Áö ÈÄ 1ºÐ °£ centrifuge. ¨èDNA polymerase, Template DNA, primers, dNTPs, mixture, reaction buffer, PCR tube, Áõ·ù¼ö, pipette tip, pipette (ÀϹÝÀûÀ¸·Î PCR ¹ÝÀÀÀº total volume 50¥ìl ·Î ¼öÇàÇÔ) 1. PCR tube¿¡ NTPs buffer 4§¡¿Í reaction buffer 5§¡¸¦ ³Ö´Â´Ù. 2. Template DNA 1§¡¸¦ ³Ö°í forward, reverse primer¸¦ °¢°¢ > |
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