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Home
>
ÀüÁ¦Ç°º¸±â
>
mRNA ¹é½Å Ä¡·áÁ¦ °³¹ß
>
in vitro transcription
> BspQ I
IVT Àû¿ë¿¡ ÃÖÀû, Type IIS Á¦ÇÑÈ¿¼Ò
BspQ I
Á¦Á¶»ç
Á¦Ç°ÄÚµå
Á¦Ç°¸í
¿ë·®
°¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í
»ç¿ëÀڸŴº¾ó
Takara
1227A
BspQ I
500 U
112,000¿ø
140,000¿ø
09.02 ~ 10.31
Takara
1227B
BspQ I
2,500 U (1227A x 5)
625,000¿ø
Product index
in vitro transcription
RUO/HQ/GMP±îÁö Ç°Áúµî..
Takara IVTpro¢â mRNA S..
PrimeCap¢â T7 RNA Poly..
Template Vector (BspQ ..
Vaccinia Capping Enzym..
mRNA Cap 2-O-Methyltra..
Faustovirus Capping En..
Takara Stable Competen..
T7 RNA Polymerase ver...
T7 RNA Polymerase
SP6 RNA Polymerase
Recombinant RNase Inhi..
Ribonucleoside 5£§-Tri..
Pyrophosphatase (inorg..
Poly(A) Polymerase
BspQ I
0.5-10 kb ssRNA Ladder..
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.
ÁÖÀÇ»çÇ×
- È¿¼ÒÀÇ °ÇÑ ±³¹ÝÀº »ï°¡ÇÑ´Ù.
¿ ½ÇÈ° Á¶°Ç
70¡É, 20ºÐ
¸ÞÆ¿ÈÀÇ ¿µÇâ
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 :
in vitro transcription
RUO/HQ/GMP±îÁö Ç°Áúµî±ÞÀ̶õ.
°íÇ°Áú °í¼öÀ² in vitro transcriptionÀ» À§ÇÑ ..
Takara IVTpro¢â mRNA Synthesis System
dsRNA »ý¼ºÀÌ Àû¾î ÀÓ»óºÐ¾ßÀÇ ¿¬±¸°³¹ß¿¡ ÀûÇÕ..
PrimeCap¢â T7 RNA Polymerase (low dsRNA)
Poly(A) ÀÌÈÄ Ãß°¡¼¿ÀÌ ¾ø´Â scarless mRNA ..
Template Vector (BspQ I) for T7 mRNA Synthesis
RNAÀÇ 5¡¯-¸»´Ü¿¡ Cap (Cap 0)À» ºÎ°¡ÇÏ´Â È¿¼Ò
Vaccinia Capping Enzyme
RNAÀÇ 5¡¯ ¸»´ÜÀ» Cap 0 ¡æ Cap 1·Î º¯È¯ÇÏ´Â ..
mRNA Cap 2-O-Methyltransferase
´Ù¾çÇÑ ¿Âµµ¿¡¼ ¹ÝÀÀÇÒ ¼ö ÀÖ´Â Capping enzym..
Faustovirus Capping Enzyme (S17)
Poly(A) ¹× ¹Ýº¹ ¼¿À» Æ÷ÇÔÇÏ´Â À¯ÀüÀÚÀÇ Å¬..
Takara Stable Competent Cells
RNA ÇÕ¼º ¼öÀ²ÀÌ Çâ»óµÈ
T7 RNA Polymerase ver.2.0
Cloned
T7 RNA Polymerase
Cloned
SP6 RNA Polymerase
Àú³óµµ DTT¿¡¼µµ ¾ÈÁ¤ÀûÀÎ RNase inhibitor
Recombinant RNase Inhibitor ver.2.0
Ribonucleoside 5£§-Triphosphates
DNA, RNA ÇÕ¼º ¹ÝÀÀÀ» ÃËÁø
Pyrophosphatase (inorganic)
Poly(A) Polymerase
IVT Àû¿ë¿¡ ÃÖÀû, Type IIS Á¦ÇÑÈ¿¼Ò
BspQ I
´Ù¾çÇÑ Å©±âÀÇ ssRNA »çÀÌÁî È®ÀÎ
0.5-10 kb ssRNA Ladder Marker