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Á¦ÇÑÈ¿¼Ò ¡¤ Quickcut
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Á¦ÇÑÈ¿¼Ò (B)
> BspQ I
IVT Àû¿ë¿¡ ÃÖÀû, Type IIS Á¦ÇÑÈ¿¼Ò
BspQ I
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(ºÎ°¡¼¼º°µµ)
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Takara
1227A
BspQ I
500 U
112,000¿ø
140,000¿ø
02.26 ~ 04.19
Takara
1227B
BspQ I
2,500 U (1227A x 5)
625,000¿ø
Product index
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Á¦ÇÑÈ¿¼Ò È°¼º¿¡ ´ëÇÑ M..
Á¦ÇÑÈ¿¼Ò Genome DNA An..
Á¦ÇÑÈ¿¼ÒÀÇ »ç¿ë¿¡ °üÇÏ..
Á¦ÇÑÈ¿¼ÒÀÇ Star È°¼º
Basal Buffer Á¶¼º
Basal BufferÀÇ ¿°³óµµ..
Double Digestion¿ë Ãß..
Ligation-Recutting È¿..
pBR322¡¢pUC19ÀÇ ¿ÏÀüºÐ..
pKF3 Cloning Test
pKF3ÀÇ ¿ÏÀüºÐÇØ¿¡ ÇÊ¿ä..
Universal Buffer ¡¤ Ba..
Á¦ÇÑÈ¿¼Ò (A)
Aat II
Acc I
Acc II £¨FnuD II£©
Acc III £¨BspM II£©
Afa I £¨Rsa I£©
Afl II
Alu I
Aor13H I £¨BspM II, Ac..
Aor51H I £¨Eco47 III£©
Apa I
ApaL I
Á¦ÇÑÈ¿¼Ò (B)
Bal I
BamH I
Ban II £¨HgiJ II£©
BciT130 I
Bcn I £¨Cau II£©
Bgl I
Bgl II
Bln I £¨Avr II£©
BmeT110 I £¨Ava I£©
Bpu1102 I £¨Esp I£©
Bsp1286 I £¨Sdu I£©
Bsp1407 I
BspQ I
BspT104 I £¨Asu II, Ns..
BspT107 I £¨HgiC I£©
BssH II £¨BseP I£©
BstP I £¨BstE II, EcoO..
BstX I
Á¦ÇÑÈ¿¼Ò (C, D)
Cfr10 I
Cla I
Cpo I £¨Rsr II£©
Dde I
Dpn I
Dra I £¨Aha III£©
Á¦ÇÑÈ¿¼Ò (E, F)
Eae I £¨Cfr I£©
Eco52 I £¨Xma III£©
Eco81 I £¨Sau I£©
EcoO109 I £¨Dra II£©
EcoO65 I £¨BstE II, Bs..
EcoR I
EcoR V
EcoT14 I £¨Sty I£©
EcoT22 I £¨Ava III£©
Fba I £¨Bcl I£©
Fok I
Á¦ÇÑÈ¿¼Ò (H, K)
Hae II
Hae III
Hap II £¨Hpa II, Msp I..
Hha I
Hinc II £¨Hind II£©
Hind III
Hinf I
Hpa I
Kpn I
Á¦ÇÑÈ¿¼Ò (M, N)
Mbo I £¨Sau3A I£©
Mbo II
Mfl I £¨Xho II£©
Mlu I
Msp I £¨Hap II, Hpa II..
Mun I £¨Mfe I£©
Nae I
Nco I
Nde I
Nhe I
Not I
Nru I
Á¦ÇÑÈ¿¼Ò (P, R)
PmaC I
Psp1406 I £¨Acl I£©
Pst I
Pvu I
Pvu II
RspRS II (Mse I)
Á¦ÇÑÈ¿¼Ò (S, T)
Sac I
Sac II
Sal I
Sau3A I £¨Mbo I£©
Sca I
Sfi I
Sma I
Smi I £¨Swa I£©
SnaB I
Spe I
Sph I
Sse8387 I
Ssp I
Stu I
Taq I £¨TthHB8 I£©
Á¦ÇÑÈ¿¼Ò (V, X)
Van91 I (PflM I)
VpaK11B I £¨Ava II£©
Xba I
Xho I
Xsp I£¨Bfa I, Mae I£©
RNAÁ¦ÇÑÈ¿¼Ò (mRNA Interfer..
mRNA Interferase¢â -Ma..
Á¦ÇÑÈ¿¼Ò QuickCut
QuickCut¢â Restriction..
QuickCut (A)
QuickCut¢â Afa I (Rsa ..
QuickCut¢â Afl II
QuickCut¢â Alu I
QuickCut¢â Apa I
QuickCut (B)
QuickCut¢â BamH I
QuickCut¢â Bgl II
QuickCut¢â Bln I (Avr ..
QuickCut (C)
QuickCut¢â Cla I
QuickCut (D)
QuickCut¢â Dpn I
QuickCut¢â Dra I (Aha ..
QuickCut (E)
QuickCut¢â EcoR I
QuickCut¢â EcoR V
QuickCut (H)
QuickCut¢â Hae III
QuickCut¢â Hha I
QuickCut¢â Hind III
QuickCut¢â Hinf I
QuickCut¢â Hpa I
QuickCut (K)
QuickCut¢â Kpn I
QuickCut (M)
QuickCut¢â Mlu I
QuickCut (N)
QuickCut¢â Nco I
QuickCut¢â Nde I
QuickCut¢â Nhe I
QuickCut¢â Not I
QuickCut (P)
QuickCut¢â Pst I
QuickCut¢â Pvu I
QuickCut¢â Pvu II
QuickCut (S)
QuickCut¢â Sac I
QuickCut¢â Sac II
QuickCut¢â Sal I
QuickCut¢â Sfi I
QuickCut¢â Sma I
QuickCut¢â SnaB I
QuickCut¢â Spe I
QuickCut¢â Sph I
QuickCut¢â Stu I
QuickCut (X)
QuickCut¢â Xba I
QuickCut¢â Xho I
Á¦ÇÑÈ¿¼Ò Àνļ¿ ÀÏÁ¤ ¿°±â ¹Ù±ù DNA¸¦ Àý´ÜÇÏ´Â Type IIS Á¦ÇÑÈ¿¼Ò (Isochizomer)
ºñ´ëĪÀÇ ÀÌÁß°¡´Ú DNA¼¿ ÀνÄ
Translation È¿À²À» ³ôÀ̱â À§ÇØ
in vitro
transcription½Ã PolyA ¼¿ ÀÌ¿Ü¿¡ ºÒÇÊ¿äÇÑ ¿°±â°¡ ¾ø´Â ÁÖÇü DNA¸¦ Á¦ÀÛÇÒ ¶§ È°¿ë
Á¦Ç°¼³¸í
Bsp
Q IÀº ºñ´ëĪÀÇ Æ¯Á¤ ÀÌÁß°¡´Ú DNA¼¿À» ÀνÄÇÏ¿© ÀÏÁ¤ ¿°±â ¹Ù±ùÀÇ DNA¸¦ Àý´ÜÇÏ´Â Type IIS Á¦ÇÑÈ¿¼ÒÀÌ´Ù (Isochizomer).
i
n vitro
transcription½Ã, mRNA ÇÕ¼ºÀ» À§ÇÑ ÁÖÇü plasmid¿¡
Bsp
Q I Àνļ¿À» Ãß°¡ÇÑ ÈÄ,
Bsp
Q IÀ» ó¸®Çϸé PolyA ¼¿ ÀÌ¿Ü¿¡ ºÒÇÊ¿äÇÑ ¿°±â°¡ ¾ø´Â mRNA¸¦ ÇÕ¼ºÇÒ ¼ö ÀÖ´Ù.
*1
*1
mRNA ÇÕ¼ºÀ» À§ÇÑ ÁÖÇü DNA¸¦ Á¦ÀÛÇÒ ¶§ PolyA ¼¿ µÚ¿¡ Ãß°¡ ¿°±â°¡ ³²´Â °æ¿ì mRNAÀÇ translation È¿À²ÀÌ ÀúÇϵǹǷΠ¿°±â°¡ ³²Áö ¾Êµµ·Ï ¼³°èÇÏ´Â °ÍÀÌ Áß¿äÇÏ´Ù (±×¸² 1).
±×¸² 1. Poly A ¼¿ µÚ¿¡ Ãß°¡µÇ´Â ¿©ºÐÀÇ ¿°±â ¼ö¿¡ µû¸¥ mRNA translation È¿À² ºñ±³
3 Á¾·ùÀÇ Á¦ÇÑÈ¿¼Ò (
Bam
H I,
Hin
d III,
Bsp
Q I)·Î ¼±ÇüÈµÈ IVT¿ë plasmid·Î mRNA ÇÕ¼º ÈÄ
Trans
IT
®
-mRNA Transfection Reagent (Code MIR 2250)
¸¦ »ç¿ëÇØ HEK293T ¼¼Æ÷¿¡ transfection ÇÏ¿´´Ù. ±× °á°ú, FLuc positive control°ú ºñ±³Çؼ 23°³ÀÇ Ãß°¡ ¿°±â°¡ ³²¾ÆÀÖ´Â mRNA´Â translation È¿À²ÀÌ Å©°Ô ÀúÇϵǾú´Ù.
¹Ý¸é¿¡ 5°³ ÀÌÇÏÀÇ ¿°±â°¡ ³²Àº °æ¿ì ¸ðµÎ ºñ½ÁÇÑ translation È¿À²À» ³ªÅ¸³ÂÁö¸¸, ¿©ºÐÀÇ ¿°±â°¡ ÀûÀ»¼ö·Ï mRNAÀÇ translation È¿À²ÀÌ ´õ ³ôÀº °æÇâÀ» º¸¿´´Ù. Ãß°¡ ¿°±â°¡ ¾øÀ» ¶§ translation È¿À²ÀÌ °¡Àå ³ô±â¿¡ mRNA ÇÕ¼ºÀ» ¸ñÀûÀ¸·Î ÇÏ´Â ÁÖÇü Á¦ÀÛ ½Ã
Bsp
Q I°ú °°Àº Type IIS Á¦ÇÑ È¿¼ÒÀÇ »ç¿ëÀ» ±ÇÀåÇÑ´Ù.
¡Ø
Hin
d IIIÁ¦ÇÑÈ¿¼Ò¸¦ »ç¿ëÇØ 3°³ ÀÌÇÏÀÇ ÀÜ¿© ¿°±â°¡ ³²´Â °æ¿ì, translation È¿À²ÀÌ ¼ÒÆø °¨¼Ò µÇ¾úÁö¸¸ ¿©ºÐ ¿°±â°¡ ¾ø´Â °æ¿ì¿Í Å« Â÷À̸¦ º¸ÀÌÁö´Â ¾Ê´Â´Ù.
¿ëµµ
mRNA ÇÕ¼º (
in vitro
transcription)¿¡ »ç¿ëµÇ´Â ÁÖÇü plasmidÀÇ ¼±ÇüÈ
³»¿ë
Bsp
Q I (10 U/§¡)
500 U
10x
Bsp
Q I Buffer
*
1 §¢
*
º» ¹öÆÛ¿¡´Â BSA°¡ Æ÷ÇԵǾî ÀÖÁö ¾Ê´Ù.
º¸Á¸ -20¡É
±â¿ø
Escherichia coli
carrying the plasmid containing the gene for
Bsp
Q I
ÀϹÝÀûÀÎ ¹ÝÀÀ Á¶°Ç
¹ÝÀÀ ¿Âµµ
50¡É
*
*
ÁÖÀÇ: 37¡É¿¡ ¹ÝÀÀ ½Ã 50¡Éº¸´Ù Àý´Ü È¿À²ÀÌ 60% Á¤µµ·Î ³·¾ÆÁø´Ù.
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- È¿¼ÒÀÇ °ÇÑ ±³¹ÝÀº »ï°¡ÇÑ´Ù.
¿ ½ÇÈ° Á¶°Ç
70¡É, 20ºÐ
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dam
methylation,
dcm
methylation¹× CpG methylationÀÇ ¿µÇâÀ» ¹ÞÁö ¾Ê´Â´Ù.
Star È°¼º
Àå½Ã°£ (e.g. 4½Ã°£ ÀÌ»ó) ¹ÝÀÀÇÒ ¶§, star È°¼ºÀÌ »ý±æ ¼ö ÀÖ´Ù.
°ü·ÃÁ¦Ç°
- Çѹø¿¡ 200 §¶ mRNA ÇÕ¼º, °í¼öÀ²
in vitro
transcription:
Takara IVTpro™ T7 mRNA Synthesis Kit (Code 6144)
*
- Uncapped RNAÀÇ 5’ ¸»´Ü¿¡ Cap 0 ±¸Á¶¸¦ ºÎ¿©ÇÏ´Â È¿¼Ò
Vaccinia Capping Enzyme (Code 2460A)
- Cap 0 ±¸Á¶¸¦ °¡Áø RNA¸¦ Cap 1À¸·Î ¸¸µå´Â È¿¼Ò
mRNA Cap 2'-O-Methyltransferase (Code 2470A)
- mRNA Àü¿ë Transfection ½Ã¾à,
Trans
IT
®
-mRNA Transfection Kit (Code MIR 2250)
- Àú³óµµ DTT¿¡¼µµ ¾ÈÁ¤ÀûÀÎ ¾÷±×·¹À̵å RNase inhibitor:
Recombinant RNase Inhibitor ver.2.0 (Code 2315A)
- RNA ÇÕ¼º ¼öÀ²ÀÌ Çâ»óµÈ Ver.2
T7 RNA Polymerase ver.2.0 (Code 2541A)
- RNA, DNA ÇÕ¼º ½Ã PPi¸¦ Á¦°ÅÇÏ¿© ¼öÀ²À» ³ôÀÌ´Â
Pyrophosphatase (inorganic) (Code 2450A)
-
Bsp
Q I°ú °°ÀÌ »ç¿ëÇÒ ¼ö ÀÖ´Â IVT¿ë Template Vector:
Template Vector (
Bsp
Q I) for T7 mRNA Synthesis (Code 6146)
*
Cloning Kit for mRNA Template (Code 6143A)
¿¡ Æ÷ÇÔµÈ IVT¿ë ÁÖÇü plasmid´Â
Bsp
Q I Á¦ÇÑÈ¿¼ÒÀÇ Àνļ¿ÀÌ Æ÷ÇԵǾî ÀÖÁö ¾Ê¾Æ »ç¿ëÀÌ ºÒ°¡´ÉÇÏ´Ù. ÀÌ Á¦Ç°À» »ç¿ëÇÒ °æ¿ì¿¡´Â
Hin
d III (Code 1060A)
¸¦ ÃßõÇÑ´Ù.
Keyword :
Á¦ÇÑÈ¿¼Ò (B)
Bal I
BamH I
Ban II £¨HgiJ II£©
BciT130 I
Bcn I £¨Cau II£©
Bgl I
Bgl II
Bln I £¨Avr II£©
BmeT110 I £¨Ava I£©
Bpu1102 I £¨Esp I£©
Bsp1286 I £¨Sdu I£©
Bsp1407 I
IVT Àû¿ë¿¡ ÃÖÀû, Type IIS Á¦ÇÑÈ¿¼Ò
BspQ I
BspT104 I £¨Asu II, Nsp V£©
BspT107 I £¨HgiC I£©
BssH II £¨BseP I£©
BstP I £¨BstE II, EcoO65 I£©
BstX I