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°íÈ¿À²ÀÇ Ligation premix ¿Í T-vectorÀÇ Á¶ÇÕ

Mighty TA-cloning Kit

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
6028
Mighty TA-cloning Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
20 ȸ
178,000¿ø  6028_e.v1812Da.pdf

Á¦Ç°¼³¸í
º» Á¦Ç°Àº Taq ±â¹ÝÀÇ DNA polymerase·Î PRCÇÑ »ê¹°ÀÇ TA-cloningÀ» À§ÇÑ Á¦Ç°ÀÌ´Ù. °íÈ¿À² ligation premixÀÎ DNA Ligation Kit £¼Mighty Mix£¾ (Code: 6023)¿Í pUC °è¿­ÀÇ pMD20-T vector¸¦ Æ÷ÇÔÇÏ°í ÀÖ¾î ºü¸£°í °£ÆíÇÏ°Ô ligationÇÒ ¼ö ÀÖ´Ù.
*TA cloning: Taq DNA polymeraseÀÇ PCR »ê¹°Àº ±× 3¡¯ ¸»´Ü¿¡ deoxyriboadenosine (dA) ÇÑ ¿°±â°¡ Ãß°¡µÈ´Ù. ÀÌ PCR ÁõÆø»ê¹°Àº 3¡¯ ¸»´Ü¿¡ deoxyribothymidine (dT) ÇÑ°³°¡ Ãß°¡µÈ T vector°¡ PCR ÁõÆø»ê¹°ÀÇ dA ¿°±â¿Í »óº¸ÀûÀ¸·Î °áÇÕÇÏ´Â °ÍÀ» ÀÌ¿ëÇØ cloningÇÒ ¼ö ÀÖ´Ù.

[ÁÖÀÇ] PrimeSTAR¢ç ½Ã¸®ÁîÀÇ PCR »ê¹°Àº ´ëºÎºÐ ÆòÈ° ¸»´Ü (blunt end)ÀÌ µÇ±â ¶§¹®¿¡ ¸»´Ü¿¡ dA ¿°±â¸¦ 1°³ºÎ°¡ÇÏ´Â °úÁ¤À» º°µµ·Î ÁøÇàÇؾ߸¸ ÀÌ Á¦Ç°À» »ç¿ëÇÒ ¼ö ÀÖ´Ù. µû¶ó¼­ ÀÌ °úÁ¤ÀÇ ½Ã¾àÀ» Æ÷ÇÔÇÑ Mighty TA-cloning Reagent Set for PrimeSTAR¢ç (Code 6019)¸¦ ÃßõÇÑ´Ù.
³»¿ë (20 ȸ)

pMD20-T vector (50 ng/§¡)

20 §¡

Ligation Mighty Mix *1

2 X 50 §¡

Positive Control Insert *2

10 §¡

*1DNA Ligation Kit £¼Mighty Mix£¾(Code 6023)¿¡ Æ÷ÇÔµÈ ½Ã¾à°ú µ¿ÀÏ
*2 3¡¯ ¸»´Ü¿¡ dA overhangÀ» °¡Áö´Â ¾à 200 bpÀÇ DNA fragment
(E.coli gDNA¸¦ ÁÖÇüÀ¸·Î TaKaRa Ex Taq¢ç¿¡¼­ ÁõÆø, 10 ng/§¡)
º¸Á¸ -20¡É
»ç¿ë»óÀÇ ÁÖÀÇ
1. Ligation Mighty Mix´Â ¾óÀ½¿¡¼­ õõÈ÷ ³ìÀÎ µÚ »ç¿ëÀü °¡º±°Ô ¼¯¾îÁØ´Ù.
2. Transformation ÈÄ¿¡µµ cloningÇÑ insert¿¡ ±æÀ̳ª ¹æÇâ¿¡ µû¶ó LB + Amp/X-Gal/IPTG plates¿¡¼­ ÆĶõ»ö colony°¡ Çü¼ºµÉ ¼ö ÀÖ´Ù. ÀÌ °æ¿ì colony PCRÀ» ÅëÇØ insert DNAÀÇ ¿©ºÎ¸¦ È®ÀÎÇÑ´Ù.
3. Agarose gel¿¡¼­ ¸ñÀû PCR »ê¹°À» ȸ¼öÇÏ´Â °æ¿ì UV¿¡ ¿À·¡ Á¶»çµÇ¸é DNA°¡ ¼Õ»óµÇ¾î cloning È¿À²ÀÌ ³·¾ÆÁú ¼ö ÀÖ´Ù.
4. PCR templateÀ¸·Î clone vector¿Í µ¿ÀÏÇÑ selection marker (pMD20-T vectorÀÇ °æ¿ì Ampr)¸¦ °¡Áø plasmid¸¦ »ç¿ëÇÏ´Â °æ¿ì, template plasmid°¡ E.coli¿¡ transformation µÇ´Â °ÍÀ» ¹æÁöÇϱâ À§ÇØ Àü±â¿µµ¿ ÈÄ agarose gel¿¡¼­ ȸ¼ö ¹× Á¤Á¦ÇÑ´Ù.
pMD20-T vector
°ü·ÃÁ¦Ç°
- PrimeSTAR¢ç µî High fidelity È¿¼Ò¸¦ ÁõÆø»ê¹°ÀÇ TA-cloning Mighty TA-cloning Reagent Set for PrimeSTAR¢ç (Code 6019)
- È¿À² ³ôÀº TA-cloningÀ» À§ÇÑ T-vector T-Vector pMD20 (Code 3270)
Âü°íÀÚ·á
- Cloning ½ÇÇè ÀÔ¹®ÀÚ¸¦ À§ÇÑ ¡®Step by Step Cloning °¡À̵塯
- ½ÇÇè ¸ñÀûº°·Î ÃßõÇÏ´Â Cloning Kit

Keyword : T4 DNA ligase,T vector,TA cloning,TA-cloning,t cloining,t-cloning,cloning,¶óÀÌ°ÔÀÌÁî,Ŭ·Î´×,t easy

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