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TaKaRa
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Cloned

Terminal Deoxynucleotidyl Transferase

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
2230A
Terminal Deoxynucleotidyl Transferase
°ü·ÃÇмú ±¸¸ÅÇϱâ
300 U
60,800¿ø 
76,000¿ø
°¡°ÝÇÒÀÎ
03.02 ~ 04.30
2230A_DS.v1901Da.pdf
Takara
2230B
Terminal Deoxynucleotidyl Transferase
°ü·ÃÇмú ±¸¸ÅÇϱâ
1,500 U
342,000¿ø 

³óµµ 7~15 U/§¡
Çü»ó
60 mM Potassium Phosphate (pH7.2)
150 mM KCl
1 mM 2-Mercaptoethanol
50% Glycerol
º¸Á¸ -20¡É
÷ºÎ Buffer Á¶¼º (5¡¿)
500 mM HEPES (pH7.2)
40 mM MgCl2
0.5 mM DTT
0.1 % BSA (º°Ã·)
Á¦Ç°¼³¸í
º» È¿¼Ò´Â ¹ÝÀÀ¿¡ ÁÖÇüÀ» ÇÊ¿ä·Î ÇÏÁö ¾Ê°í, ´ÜÀϰ¡´Ú ¶Ç´Â ÀÌÁß°¡´Ú DNAÀÇ 3¡¯- OH ¸»´Ü¿¡ deoxynucleotide¸¦ ÁßÇÕÇÏ´Â ¹ÝÀÀÀ» Ã˸ÅÇÑ´Ù. ÇÁ¶óÀ̸Ӱ¡ µÇ´Â ÃÖÀú 3¿°±â ÀÌ»óÀÇ oligonucleotide°¡ ÇÊ¿äÇÏ´Ù.

TdT¸¦ ÀÌ¿ëÇÑ ´ÜÀÏ ÁßÇÕüÀÇ ºÎ°¡ ¹ÝÀÀÀÇ Á¶°ÇÀº
   1. ºÎ°¡ÇÏ´Â ¿°±âÀÇ Á¾·ù (dATP, dTTP, dGTP, dCTP)
   2. ¹öÆÛ ÁßÀÇ 2°¡ ÀÌ¿ÂÀÇ Á¾·ù (Mn2+, Co2+, Mg2+)
   3. ºÎ°¡µÇ´Â DNAÀÇ ¸»´Ü±¸Á¶ (µ¹Ãâ 3¡¯- OH¸»´Ü, ÆòȰ¸»´Ü, ÇÔ¸ô 3¡¯- OH¸»´Ü) µîÀÇ ¿µÇâÀ» ¹Þ±â ¶§¹®¿¡ »óȲ¿¡ µû¶ó ÃÖÀûÀÇ Á¶°ÇÀ» ã¾Æ¾ß ÇÑ´Ù.
Ȱ¼ºÀÇ Á¤ÀÇ
DNase ó¸® ÈÄ ¿­º¯¼º ¼Û¾ÆÁö Èä¼± DNA (Ȱ¼ºÈ­ ¼Û¾ÆÁö Èä¼± DNA)¸¦ initiator·Î »ç¿ëÇÏ¿© 37¡É, pH7.2 Á¶°Ç¿¡¼­ 1½Ã°£µ¿¾È ¸ðµç 1 nmolÀÇ [3H]dATP¸¦ »êºÒ¿ë¼º ħÀü¹°ÀÇ ÇÕ¼º±âÁú·Î ¹Ù²Ù´Â È¿¼ÒȰ¼º 1 UÀ¸·Î ÇÑ´Ù.
Ȱ¼º ÃøÁ¤¿ë ¹ÝÀÀ¾× Á¶¼º
67 mM Tris-HCl (pH8.8)
6.7 mM MgCl2
16.6 mM (NH4)2SO4
10 mM 2-Mercaptoethanol
6.7 ¥ìM EDTA
0.0167% BSA
0.2 mg/ml ±âÁú DNA
°¢ 330 ¥ìM dCTP, dATP, dGTP
330 ¥ìM [3H]dTTP
¼øµµ
- 15 UÀÇ º» È¿¼Ò¿Í 1§¶ÀÇ ¥ëDNA-Hind III ºÐÇØ¹°À» 37¡É, 16½Ã°£ ¹ÝÀÀÇÏ¿©µµ DNAÀÇ Àü±â¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ¾ø´Ù.
- 15 UÀÇ º» È¿¼Ò¿Í 1§¶ÀÇ closed circular (RF I) pBR322 DNA¸¦ 37¡É, 16½Ã°£ ¹ÝÀÀÇÏ¿©µµ DNAÀÇ Àü±â¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ¾ø´Ù.
- 15 UÀÇ º» È¿¼Ò¿Í 1§¶ÀÇ 16S ¹× 23S RNA¸¦ 37¡É, 5½Ã°£ ¹ÝÀÀÇÏ¿©µµ RNA ÀÇ Àü±â ¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ¾ø´Ù.
¿ëµµ
- Okayama-Berg¹ý¿¡ ÀÇÇÑ º¤ÅÍ-cDNA¿¡ »óº¸ÀûÀÎ ´ÜÀÏ ÁßÇÔüÀÇ ºÎ°¡
- Ç¥½Ä dNTP ¶Ç´Â ddNTP¿¡ ÀÇÇÑ DNAÀÇ 3¡¯¸»´ÜÇ¥½Ä

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