Mouse Tail Direct PCR Kit
|
|
|
|
|
Mouse tail·Î ºÎÅÍ DNA purification °úÁ¤À» °ÅÄ¡Áö ¾Ê°í ¹Ù·Î PCRÀ» ÁøÇàÇÒ ¼ö ÀÖÀ¸¹Ç·Î mouse tailÀ» ÀÌ¿ëÇÑ genotyping¿¡ Æí¸®ÇÑ Á¦Ç°ÀÔ´Ï´Ù. |
|
|
KOMA |
ÇöÀç Á¦Ç°°ü·Ã ¹®ÀÇÇϱâ
ÀμâÇϱâ
- Transgeniec mouse genotyping¿¡ È¿°úÀûÀÔ´Ï´Ù.
- DNA¸¦ »ÌÁö¾Ê°í mouse tail·Î ¹Ù·Î PCRÀ» ÇÒ ¼ö ÀÖ¾î ½Ã°£ÀÌ ´ÜÃàµÇ¸ç Æí¸®ÇÕ´Ï´Ù.
- Proteinase K (PK)¸¦ ó¸®ÇÒ Çʿ䰡 ¾øÀ¸¸ç, overnight 󸮾øÀÌ, ´Ü 10ºÐ incubationÀ» ÅëÇØ template¸¦ ¾òÀ» ¼ö ÀÖ½À´Ï´Ù.
- Hot Start TaqÀ» ÀÌ¿ëÇÏ¿© ºñƯÀÌ ¹ÝÀÀÀ» ÁÙÀ̰í PCR È¿À²À» ³ôÇûÀ¸¸ç, Direct PCR Á¶°ÇÀ» ÃÖÀûÈ ÇÏ¿´½À´Ï´Ù.
- ½ÇÇè°úÁ¤ÀÌ ´Ü¼øÇÕ´Ï´Ù.
- 1) Mouse tailÀ» 1-2mm Å©±â·Î ÀÚ¸¥´Ù.
- 2) ÇÔ²² Á¦°øµÇ´Â Lysis buffer¿¡ ³Ö°í 60¡ÆC¿¡¼ 10ºÐ°£ incubation ÇÑ´Ù.
- 3) Vortex³ª spin down
- 4) 1ul¸¦ ¶°¼ ¹Ù·Î PCRÀ» ¼öÇàÇÑ´Ù.

Á¦Ç°ÀÇ ±¸¼º
| 2X Direct Hot Taq PCR MasterMix |
|
1ml |
| 5X Direct Lysis Buffer |
|
1ml |
| Magic Buffer |
|
1.5ml |

»ç¿ë¿¹
Direct PCR from mouse tail tissue :
- 10mgÀÇ mouse tailÀ» lysis buffer·Î lysis¸¦ ¼öÇàÇÏ¿© °¢°¢ 2, 0.2, 0.04, 0.02ul ÀÇ ³óµµ·Î PCRÀ» ¼öÇàÇÏ¿´´Ù. PK 󸮾øÀÌ 60¡ÆC¿¡¼ 10ºÐ incubation ¸¸À¸·Îµµ ±ú²ýÇÏ°Ô PCR ÁõÆøÀÌ ÀÌ·ç¾îÁüÀ» È®ÀÎÇÒ¼ö ÀÖ¾ú´Ù.
- Lane 1,3,5,7 : Direct PCR after lysis using Proteinase K at 56¡ÆC for 10 min and 95¡ÆC for 5 min
- Lane 2,4,6,8: Direct PCR without PK treatment. Incubation at 60¡ÆC for 10 min