» ÀÛ¼ºÀÚ : KSMCB | » ÀÛ¼ºÀÏ : 2018-11-27 | » Á¶È¸ : 1488 |
» ÷ºÎÆÄÀÏ : | ÷ºÎÆÄÀÏÀÌ ¾ø½À´Ï´Ù. |
°í¸¶¹ÙÀÌ¿ÀÅØ(ÁÖ) - Mouse Tail Direct PCR Kit Mouse tail·Î ºÎÅÍ DNA ÃßÃâ¾øÀÌ 10ºÐ¾È¿¡ ¹Ù·Î PCRÀ» ÁøÇàÇÒ ¼ö ÀÖ´Â kit ÀÔ´Ï´Ù. ¡à DNA¸¦ »ÌÁö¾Ê°í mouse tail·Î ¹Ù·Î PCRÀ» ¼öÇàÇϹǷÎ, ½Ã°£ÀÌ ´ÜÃàµÇ¸ç °£ÆíÇÕ´Ï´Ù. ¡à Proteinase K (PK)¸¦ ó¸®ÇÒ ÇÊ¿ä°¡ ¾ø¾î ºñ¿ë°ú ½ºÅÜÀ» ÁÙÀϼö ÀÖ½À´Ï´Ù. ¡à Overnight incubationÀº ±×¸¸! ´Ü 10ºÐ lysis °úÁ¤À» ÅëÇØ template¸¦ ¾òÀ» ¼ö ÀÖ½À´Ï´Ù. ¡à Hot Start TaqÀ» ÀÌ¿ëÇÏ¿© ºñƯÀÌ ¹ÝÀÀÀ» ÁÙÀÌ°í PCR È¿À²À» ³ôÇû½À´Ï´Ù. ¡à Direct PCR buffer¸¦ mouse tail Á¶°Ç¿¡ ÃÖÀûȽÃÄÑ, ÁÁÀº °á°ú¸¦ º¸ÀåÇÕ´Ï´Ù. ¡à Á¶ÀÛÀ» ÃÖ¼ÒÈÇÔÀ¸·Î½á ¿À¿°ÀÇ À§Çè°ú ¹ø°Å·Î¿òÀ» ÁÙ¿©ÁÝ´Ï´Ù. ¡à Transgenic mouse genotyping¿¡ Àû¿ëÇϸé È¿°úÀûÀÔ´Ï´Ù. ¡à Mouse ear·Îµµ »ç¿ëÀÌ °¡´ÉÇÕ´Ï´Ù.
1) Mouse tailÀ» 1-2mm Å©±â·Î ÀÚ¸¥´Ù. 2) ÇÔ²² Á¦°øµÇ´Â Lysis buffer¿¡ ³Ö°í 60°C¿¡¼ 10ºÐ°£ incubation ÇÑ´Ù. 3) Vortex³ª spin down 4) »óÃþ¾× 1ul¸¦ ¶°¼ ¹Ù·Î PCRÀ» ¼öÇàÇÑ´Ù. Direct PCR from mouse tail tissue : o 10mgÀÇ mouse tailÀ» lysis buffer·Î lysis¸¦ ¼öÇàÇÏ¿© °¢°¢ 2, 0.2, 0.04, 0.02ul ÀÇ ³óµµ·Î PCRÀ» ¼öÇàÇÏ¿´´Ù. PK 󸮾øÀÌ 60°C¿¡¼ 10ºÐ incubation ¸¸À¸·Îµµ ±ú²ýÇÏ°Ô PCR ÁõÆøÀÌ ÀÌ·ç¾îÁüÀ» È®ÀÎÇÒ¼ö ÀÖ¾ú´Ù. o Lane 1,3,5,7 : Direct PCR after lysis using Proteinase K at 56°C for 10 min and 95°C for 5 min o Lane 2,4,6,8: Direct PCR without PK treatment. Incubation at 60°C for 10 min - ¹®ÀÇó : °í¸¶¹ÙÀÌ¿ÀÅØ¢ß Tel:02-579-8787, Fax:02-578-7042E-mail : koma@komabiotech.co.kr, Hompage : http://www.komabiotech.co.kr - ÷ºÎ : °ü·Ã ¸µÅ© |