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ºÐÀÚ»ý¹°ÇÐ - RNA ÃßÃâ ½ÇÇè ·¹Æ÷Æ®
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1. Title : RNA ÃßÃâ°ú RT-PCR, PCR
4. Materials
5. Method
[ RNA ̧̉ ]
[ RT-PCR ]
6. Result
7. ½ÇÇè ½Ã ÁÖÀÇ »çÇ×
8. RNA ÃßÃâ ¿ø¸®
9. RT-PCR
10. PCR
11. ÀÌ ¿ÜÀÇ ½Ã¾àÀ» ÷°¡ÇÏ´Â ÀÌÀ¯
12. Reference
º» ÀÚ·á´Â ¿öµð¾ÈÀ̳ª ÇÑ±Û 2002 ÀÌ»óÀÇ ¹öÀü¿¡¼¸¸ È®ÀÎÀÌ °¡´ÉÇÕ´Ï´Ù.
ÇÑ±Û 97 ÀÌÇÏÀÇ ¹öÀüÀ» º¸À¯ÇÏ°í °è½Å ȸ¿ø´Ôµé²²¼´Â ±¸¸Å¿¡ ¾Õ¼ Âü°íÇϽñ⠹ٶø´Ï´Ù.
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1. Title : RNA ÃßÃâ°ú RT-PCR, PCR
4. Materials
(1) Reagents : PBS, Trizol, chloroform, isopropyl alcohol, 70% EtOH, RNase free water, PCR Mixture, RT-PCR Mix 1, Mix 2
(2) Instruments : pipette, tip, scraper, e-tube, centrifuge, vortex, PCR ±â°è, ice, Àå°©,kimwipes
5. Method
[ RNA ̧̉ ]
¨ç ¹èÁö¸¦ ¹ö¸° ÈÄ, PBS 2ml·Î washingÀ» 2ȸÇÑ´Ù.
¨è Trizol 2ml¸¦ ÷°¡ ÈÄ scraper·Î ±Ü´Â´Ù.
¨é e-tube 2°³¿¡ 1ml¾¿ ³ª´©¾î ´ã´Â´Ù.
¨ê chloroformÀ» 300 ÷°¡ÇÑ ÈÄ vortexingÇÑ´Ù.
¨ë 12000rpm, 4¡É, 10ºÐ°£ centrifugeÇÑ´Ù. (¾Æ·¡¿Í °°Àº °á°ú°¡ ³ªÅ¸³´Ù.)
¨ì pipetteÀÇ tipÀ» Ç¥¸é °¡±îÀÌ¿¡¼ õõÈ÷ e-tube¿¡ »óµî¾×¸¸ 400 ÃëÇÑ´Ù.
¨í isopropyl alcoholÀ» »óµî¾×°ú µ¿·®ÀÎ 400 ÷°¡ÇÑ ÈÄ 10ºÐ°£ Á¤Ä¡ÇÑ´Ù.
¨î 12000rpm, 4¡É, 10ºÐ°£ centrifugeÇÑ´Ù.
¨ï »óµî¾×À» µû¶ó ¹ö¸°´Ù.
¨ð 70% EtOH 1 ÷°¡ÇÑ ÈÄ, tube¸¦ up & down ÇÑ ÈÄ, spin downÇÑ´Ù.
¨ñ 7500rpm, 4¡É, 5ºÐ°£ centrifugeÇÑ´Ù.
¨ò »óµî¾×(70% EtOH)À» tipÀ¸·Î »ì»ì ±Ü¾î³»¾î Á¦°ÅÇÑ´Ù.
¨ó kimwipes¿¡ °Å²Ù·Î ³õ¾ÆµÎ°í dryÇÑ´Ù.
¨ô RNase free water 30¿¡ ³ìÀδÙ.
¨õ 55¡É¿¡¼ 10ºÐ°£ boilingÇÑ´Ù.
⑯ PCR Mixture¸¦ À§ÇØ SpectrophotometryÀ» ÇÏ¿© RNAÀÇ ¾çÀ» °è»êÇÑ´Ù.
(Result¿¡ RNA ¾ç °è»ê)
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http://blog.naver.com/dextermh/10011206851
http://www.genehunters.co.kr/Training/01_3.htm
http://blog.naver.com/tjdndud435?Redirect=Log&logNo=100023341953
http://www.chop.edu/research/facil/Napcore/OligoCalc.htm
http://www.uct.ac.za/depts/mmi/bbhelp/intro.html
http://cafe.naver.com/takarapcr.cafe?iframe_url=/ArticleRead.nhn%3Farticleid=41
http://cafe.naver.com/takarapcr.cafe?iframe_url=/ArticleRead.nhn%3Farticleid=1
http://www.takara.co.kr/ |
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