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mRNA ¹é½Å Ä¡·áÁ¦ °³¹ß
>
in vitro transcription
> Template Vector (BspQ I) for T7 mRNA Synthesis
Poly(A) ÀÌÈÄ Ãß°¡¼¿ÀÌ ¾ø´Â scarless mRNA ÇÕ¼ºÀÌ °¡´ÉÇÑ linearized template vector
Template Vector (BspQ I) for T7 mRNA Synthesis
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(ºÎ°¡¼¼º°µµ)
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Takara
6146
Template Vector (BspQ I) for T7 mRNA Synthesis
10 §¡
87,200¿ø
109,000¿ø
09.02 ~ 10.31
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..
in vitro
transcription (IVT)¿¡¼ »ç¿ëÇÏ´Â mRNA ÇÕ¼º¿ë plasmid¸¦ Á¦ÀÛÇϱâ À§ÇÑ template
Type IIS Á¦ÇÑÈ¿¼Ò
Bsp
Q I (Code 1227A)
Àνļ¿ÀÌ ÀÖ¾î Poly (A) ÀÌÈÄ Ãß°¡¼¿ÀÌ ¾ø´Â scarless mRNA ÇÕ¼º °¡´É
In-Fusion
¢ç
Snap Assembly Master Mix (Code 638947 ¿Ü)
(º°µµ±¸¸Å)¸¦ »ç¿ëÇØ ºü¸£°í °£´ÜÇÑ cloning
Á¦Ç° ¼³¸í
ÀÌ Á¦Ç°Àº TriLinkÀÇ Cap analog (CleanCap Reagent AG ¶Ç´Â CleanCap Reagent AG (3¡Ç OMe))¸¦ »ç¿ëÇÏ¿©mRNA¸¦ ÇÕ¼ºÇÒ ¶§ ÇÔ²² »ç¿ëÇÒ ¼ö ÀÖ´Â templateÀÌ´Ù. ÀÌ linearized template vector¿¡´Â T7 promoter, transcription start sequence (AGG), 5¡¯-UTR (untranslated region), 3¡¯-UTR, Poly(A) ¼¿ (141 base)ÀÌ Æ÷ÇԵǾî ÀÖ´Ù. µû¶ó¼ ¹ßÇö½ÃÅ°°íÀÚ ÇÏ´Â À¯ÀüÀÚÀÇ CDS (coding sequence)¸¸
In-Fusion
¢ç
À» ÅëÇØ cloning Çϸé humanÀ̳ª mouse µî mammalian cell¿¡ »ç¿ë °¡´ÉÇÑ mRNA ÇÕ¼º¿ë plasmid (CleanCap Reagent AG ¿ë)¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù (±×¸² 1).
¶ÇÇÑ Poly(A) ¼¿ ¹Ù·Î ´ÙÀ½¿¡ À§Ä¡ÇÑ Type IIS Á¦ÇÑÈ¿¼Ò
Bsp
Q I
À¸·Î ¼±ÇüÈ ÇÏ¿© Poly(A) ÀÌÈÄ Ãß°¡¼¿ÀÌ ¾ø´Â mRNA ÇÕ¼º¿ë templateÀ» Á¦ÀÛÇÒ ¼ö ÀÖ´Ù (±×¸² 2). ÀÌ templateÀº
Takara IVTpro¢â T7 mRNA Synthesis Kit (Code 6144)
¸¦ »ç¿ëÇÏ¿© °£ÆíÇÏ°Ô °í¼öÀ²·Î ¸ñÀû mRNA¸¦ ÇÕ¼ºÇϴµ¥ »ç¿ëµÈ´Ù.
mRNA ÇÕ¼º¿ë plasmidÀÇ ¼³°è ¹× ÀÚ¼¼ÇÑ ÇÁ·ÎÅäÄÝÀº ÀÌ Á¦Ç°ÀÇ »ç¿ëÀڸŴº¾ó°ú ÇÔ²²
Cloning Kit for mRNA Template (Code 6143)
ÀÇ »ç¿ëÀÚ ¸Å´º¾óÀ» Âü°íÇϼ¼¿ä.
±×¸² 1. In-Fusion
¢ç
À» »ç¿ëÇÑ IVT¿ë template plasmid Á¦ÀÛ
±×¸² 2. Á¦ÇÑÈ¿¼Ò
Bsp
Q I Àνļ¿ ÁÖº¯ ¼¿ ¹× ¼±ÇüÈ ÀÌÈÄÀÇ ¸»´Ü¼¿ ±¸Á¶
- Type IIS Á¦ÇÑÈ¿¼Ò
Bsp
Q IÀÇ Àνļ¿ (»¡°»ö) ¹× Àý´ÜÀ§Ä¡ (ÆĶõ»ö)
- Type IIS Á¦ÇÑÈ¿¼ÒÀÇ Àý´ÜÀ§Ä¡´Â Àνļ¿ÀÇ ¿ÜºÎ¿¡ À§Ä¡ÇÑ´Ù. µû¶ó¼
Bsp
Q IÀ» »ç¿ëÇÏ´Â °ÍÀ¸·Î Poly(A) ÀÌÈÄ ¿©ºÐ ¼¿ÀÌ ¾ø´Â mRNA ÇÕ¼º¿ë templateÀ» Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.
¿ëµµ
Poly (A) ÀÌÈÄ Ãß°¡ ¼¿ÀÌ ¾ø´Â mRNA ÇÕ¼º¿ë template Á¦ÀÛ
³»¿ë
Linearized Template Vector (
Bsp
Q I) (50 ng/§¡)
10 §¡
FLuc Control Fragment (
Bsp
Q I) (100 ng/§¡)
10 §¡
*
¼¿ Á¤º¸´Â ¾Æ·¡¸¦ Âü°í.
¼¿ Á¤º¸
- Linearized Template Vector (BspQ I)
:
sequence fasta ÆÄÀÏ ¹Ù·Î°¡±â
- FLuc Control Fragment (BspQ I)
:
sequence fasta ÆÄÀÏ ¹Ù·Î°¡±â
º¤Å͸Ê
Neo
R
/Kan
R
53¢¦847
Ori: 1,172¢¦1,760
CMV enhancer: 1,861¢¦2,240
CMV promoter: 2,241¢¦2,444
T7 promoter sequence: 2,489¢¦2,505
Transcription start sequence (AGG): 2,506¢¦2,508
5¡¯UTR: 2,509¢¦2,559
Upstream In-Fusion region (15 bp): 2,545¢¦2,559
¡åIn-Fusion cloning site (linearized site)
: between 2,559 and 2,560
Downstream In-Fusion region (15 bp): 2,560¢¦2,574
3¡¯UTR: 2,560¢¦2,672
Poly(A)141 : 2,673¢¦2,813
Bsp
Q I: 2,810
Poly(A) Forward Primer: 2,570¢¦2,589
Poly(A) Reverse Primer: 2,904¢¦2,923
ÁÖÀÇ»çÇ×
- ÀÌ Á¦Ç°À¸·Î IVT¿ë template vector¸¦ ¸¸µé±â À§Çؼ´Â
In-Fusion
¢ç
Snap Assembly Master Mix (Code 638947 ¿Ü)
°¡ ÇÊ¿äÇÕ´Ï´Ù.
- mRNA ÇÕ¼º¿ë plasmidÀÇ ¼³°è ¹× ÇÁ·ÎÅäÄÝ¿¡ ´ëÇÑ ÀÚ¼¼ÇÑ ³»¿ëÀº ÀÌ Á¦Ç°ÀÇ »ç¿ëÀÚ ¸Þ´º¾ó°ú ÇÔ²²
Cloning Kit for mRNA Template (Code 6143)
ÀÇ »ç¿ëÀÚ ¸Å´º¾óÀ» Âü°íÇϼ¼¿ä. À̶§ CDS¿¡
Bsp
Q I Àνļ¿ÀÌ ÀÖ´ÂÁö È®ÀÎÀÌ ÇÊ¿äÇÕ´Ï´Ù. Àý´ÜºÎÀ§ ÀÌ¿Ü¿¡
Bsp
Q I Àνļ¿ÀÌ Á¸ÀçÇÏ´Â °æ¿ì, ¾Æ¹Ì³ë»ê ¼¿¿¡ º¯È°¡ »ý±âÁö ¾Ê´Â ´Ù¸¥ codon (eg. Alanine codon À» ¡°GCU¡±¿¡¼ ¡°GCC¡±·Î º¯°æ)À¸·Î º¯°æÇÏ¿© »ç¿ëÇϼ¼¿ä.
- Template plasmid¿¡ RNase°¡ È¥ÀԵǸé RNAÀÇ ¼öÀ²ÀÌ ÀúÇϵǰųª fragmentationÀÌ ¹ß»ýÇÕ´Ï´Ù. ¹ÝÀÀ¿¡ »ç¿ëÇÏ´Â Æ©ºê³ª ¸¶ÀÌÅ©·ÎÆÄÀÌÆê¿ë ÆÁÀ» RNA ÇÕ¼º Àü¿ëÀ¸·Î »ç¿ëÇÏ°í ÀÏȸ¿ë Àå°©À» Âø¿ëÇÏ¿© RNase°¡ È¥ÀÔµÇÁö ¾Êµµ·Ï ÁÖÀÇÇϼ¼¿ä.
º¸Á¸: - 20¡É
°ü·ÃÁ¦Ç°
- IVTÀü vectorÀÇ ¼±ÇüÈ¿¡ »ç¿ëÇÏ´Â Type IIS Á¦ÇÑÈ¿¼Ò
Bsp
Q I (Code 1227A)
- Çѹø¿¡ 200 §¶ mRNA ÇÕ¼º, °í¼öÀ² in vitro transcription:
Takara IVTpro¢â T7 mRNA Synthesis Kit (Code 6144)
- In-Fusion
¢ç
cloning½Ã °³¼±µÈ colony Çü¼º ´É·ÂÀ¸·Î ¼º´ÉÀº up °¡°ÝÀº downµÈ
In-Fusion
¢ç
Snap Assembly Master Mix (Code 638947)
- In-Fusion
¢ç
cloning¿¡ ÃÖÀûÈµÈ competent cell
Stellar¢â Competent Cells (Code 636763)
- ¹ü¿ë¼º ³ôÀº High fidelity PCR È¿¼Ò
PrimeSTAR
¢ç
GXL DNA Polymerase (Code R050A)
- ½ÇÇè ȯ°æ¿¡ ÀÖ´Â RNase ±ò²û Á¦°Å
RNase-OFF
¢ç
(RNase Contamination Á¦°Å ¿ë¾×) (Code 9037)
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