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ÀüÁ¦Ç°º¸±â
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Cloning °ü·Ã
>
¼ö½ÄÈ¿¼Ò£¯DNA Polymerase¡¢RNA Polymerase
> DNA Polymerase I (E. coli )
Cloned
DNA Polymerase I (E. coli )
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Takara
2130A
DNA Polymerase I£¨E. coli£©
500 U
74,000¿ø
Takara
2130B
DNA Polymerase I£¨E. coli£©
2,500 U
333,000¿ø
³óµµ 3~6 U/§¡
Çü»ó
50 mM
Potassium Phosphate (pH6.5)
10 mM
2-Mercaptoethanol
50%
Glycerol
º¸Á¸ -20¡É
÷ºÎ Buffer Á¶¼º (10¡¿)
500 mM
Tris-HCl (pH7.8)
100 mM
MgCl
2
1 mM
DTT
250 §¶/ml
BSA*
*BSA°¡ ħÀüµÉ ¿ì·Á°¡ ÀÖÀ¸¹Ç·Î µ¿°á¡¤ À¶ÇØ´Â °¡´ÉÇÑ ÇÇÇÑ´Ù.
±â¿ø
Escherichia coli carrying the plasmid which encodes the gene of Pol I
Á¦Ç°¼³¸í
º» È¿¼Ò´Â ÁÖÇü, ÇÁ¶óÀ̸Ó(DNA, RNA ¸ðµÎ °¡´É) Á¸ÀçÇÏ¿¡¼ dNTP¸¦ ±âÁú·Î ÇÏ¿©, ÁÖÇü¿¡ »óº¸ÀûÀÎ DNA¸¦ 5¡¯¡æ 3¡¯¹æÇâÀ¸·Î ÇÕ¼ºÇÑ´Ù. ÀÌÁß°¡´Ú ƯÀÌÀû 5¡¯¡æ3¡¯exonuclease Ȱ¼º ¹× ´ÜÀϰ¡´Ú ƯÀÌÀû 3¡¯¡æ 5¡¯exonuclease Ȱ¼ºµµ °¡Áø´Ù.
Ȱ¼ºÀÇ Á¤ÀÇ
Poly d(A-T)ÇÕ¼º DNA¸¦ ÁÖÇü/ÇÁ¶óÀ̸ӷΠÇÏ¿© 37¡É, pH7.4¿¡¼, 30ºÐ µ¿¾È ¸ðµç 10 nmol nucleotide¸¦ »êºÒ¿ë¼º ħÀü¹°·Î ¹Ù²Ù´Â È¿¼Ò Ȱ¼ºÀ» 1 UÀ¸·Î ÇÑ´Ù.
Ȱ¼ºÃøÁ¤¿ë ¹ÝÀÀ¾×Á¶¼º
67 mM
Potassium phosphate (pH7.4)
6.7 mM
MgCl
2
1 mM
2-Mercaptoethanol
20 ¥ìM
±âÁú DNA
33 ¥ìM
dATP
33 ¥ìM
[
3
H]dTTP
¼øµµ
- SDS Àü±â¿µµ¿¿¡¼ 90% ÀÌ»óÀÇ ¼øµµ¸¦ ³ªÅ¸³½´Ù.
- 10 UÀÇ º» È¿¼Ò¿Í 1§¶ÀÇ closed circular (RF I)¬æX174 DNA¸¦ 37¡É, 1½Ã°£ ¹ÝÀÀÇÏ¿©µµ DNAÀÇ Àü±â¿µµ¿ ÆÐÅÏ¿¡ º¯È°¡ ÀϾÁö ¾Ê´Â´Ù.
»ç¿ë»óÀÇ ÁÖÀÇ
- »ó±â º¸Á¸Á¶°Ç¿¡¼ 6°³¿ù ÀÌ»ó ¾ÈÁ¤Çϰí Èñ¼®¿¡ ÀÇÇÑ È°¼ºÀúÇÏ´Â ÀϾÁö ¾ÊÀ¸³ª, ½ÉÇÏ°Ô ±³¹ÝÇÏ¸é ½ÇȰÇÒ ¼öµµ ÀÖ´Ù.
- Endonuclease¸¦ Æ÷ÇÔÇϰí ÀÖÁö ¾ÊÀ¸¹Ç·Î ´Üµ¶À¸·Î´Â °ÅÀÇ nick translation Ȱ¼ºÀº ³ªÅ¸³»Áö ¾Ê´Â´Ù.
- DNA¿Í ģȼºÀÌ °Çϱ⠶§¹®¿¡ °ú·®À» »ç¿ëÇϸé aggregationÀÌ ÀϾ ¹ÝÀÀÀÌ ÃæºÐÈ÷ ÁøÇàµÇÁö ¾Ê´Â °æ¿ì°¡ ÀÖ´Ù.
¿ëµµ
- DNase I (TaKaRa Code 2210)°úÀÇ º´¿ë¿¡ ÀÇÇÑ nick translation
- Okayama-Berg¹ý¿¡ ÀÖ¾î¼ second strandÀÇ ÇÕ¼º¹ÝÀÀ (DNA polymerase I 0.3 §¶Àº ¾à 2.5 U¿¡ »ó´çÇÑ´Ù).
Keyword :
¼ö½ÄÈ¿¼Ò£¯DNA Polymerase¡¢RNA Polymerase
Cloned
DNA Polymerase I (E. coli )
Cloned
Klenow Fragment (Large Fragment E. coli DNA Polymerase I)
Poly(A) Polymerase
Cloned
SP6 RNA Polymerase
Cloned
T4 DNA Polymerase
Cloned
T7 RNA Polymerase
Cloned
Terminal Deoxynucleotidyl Transferase