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Home > ÀüÁ¦Ç°º¸±â > 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

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
6146
Template Vector (BspQ I) for T7 mRNA Synthesis
°ü·ÃÇмú ±¸¸ÅÇϱâ
10 §¡
87,200¿ø 
109,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
6146_DS.pdf

  • in vitro transcription (IVT)¿¡¼­ »ç¿ëÇÏ´Â mRNA ÇÕ¼º¿ë plasmid¸¦ Á¦ÀÛÇϱâ À§ÇÑ template
  • Type IIS Á¦ÇÑÈ¿¼Ò BspQ 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¢çÀ» 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 BspQ IÀ¸·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¢çÀ» »ç¿ëÇÑ IVT¿ë template plasmid Á¦ÀÛ


±×¸² 2. Á¦ÇÑÈ¿¼Ò BspQ I Àνļ­¿­ ÁÖº¯ ¼­¿­ ¹× ¼±ÇüÈ­ ÀÌÈÄÀÇ ¸»´Ü¼­¿­ ±¸Á¶
- Type IIS Á¦ÇÑÈ¿¼Ò BspQ 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 BspQ 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 (BspQ I) (50 ng/§¡)

10 §¡

FLuc Control Fragment (BspQ 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

NeoR/KanR 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
BspQ 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¿¡ BspQ I Àνļ­¿­ÀÌ ÀÖ´ÂÁö È®ÀÎÀÌ ÇÊ¿äÇÕ´Ï´Ù. Àý´ÜºÎÀ§ ÀÌ¿Ü¿¡ BspQ 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 BspQ 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
>
°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
>
Àú³óµµ DTT¿¡¼­µµ ¾ÈÁ¤ÀûÀÎ RNase inhibitor
Recombinant RNase Inhibitor ver.2.0
>
DNA, RNA ÇÕ¼º ¹ÝÀÀÀ» ÃËÁø
Pyrophosphatase (inorganic)
>
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