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Home > ÀüÁ¦Ç°º¸±â > Cloning °ü·Ã > In-Fusion Cloning > [NEW] In-Fusion¢ç Snap Assembly
ÃÖ°íÀÇ Colony Çü¼º´É°ú Á¤È®ÇÑ PCR cloning

[NEW] In-Fusion¢ç Snap Assembly

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
638947
In-Fusion¢ç Snap Assembly Master Mix
°ü·ÃÇмú ±¸¸ÅÇϱâ
10 ȸ
234,400¿ø 
293,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x111003, x111002
Clontech
638948
In-Fusion¢ç Snap Assembly Master Mix
°ü·ÃÇмú ±¸¸ÅÇϱâ
50 ȸ
995,200¿ø 
1,244,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
638949
In-Fusion¢ç Snap Assembly Master Mix
°ü·ÃÇмú ±¸¸ÅÇϱâ
250 ȸ
3,236,000¿ø 
4,045,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
638943
In-Fusion¢ç Snap Assembly Master Mix
°ü·ÃÇмú ±¸¸ÅÇϱâ
500 ȸ (638949 x 2)
5,911,200¿ø 
7,389,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
638944
In-Fusion¢ç Snap Assembly Master Mix
°ü·ÃÇмú ±¸¸ÅÇϱâ
1,000 ȸ (638949 x 4)
11,200,000¿ø 
14,000,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
639615
Cloning Enhancer
°ü·ÃÇмú ±¸¸ÅÇϱâ
100 ȸ
314,400¿ø 
393,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x342553
Clontech
638951
In-Fusion¢ç Snap Assembly Master Mix with Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
10 ȸ
(Package)
424,800¿ø 
531,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x111002, x109122, x33081
Clontech
638952
In-Fusion¢ç Snap Assembly Master Mix with Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
50 ȸ
(Package)
1,955,200¿ø 
2,444,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
636763
Stellar¢â Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
10 ȸ (10 x 100ul)
234,400¿ø 
293,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x33081
Clontech
636766
Stellar¢â Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
50 ȸ (636763 x 5)
996,800¿ø 
1,246,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
636767
Stellar¢â Competent Cells (96 well plate)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
96 ȸ (96 x 20ul)
(Package)
657,600¿ø 
822,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x33079

¡ØÂü°í»çÇ×
Ÿ»ç ´ëºñ ¶Ù¾î³­ Á¤È®µµ¸¦ °¡Áø In-Fusion¢çÀÇ ÀÚ¼¼ÇÑ Â÷ÀÌÁ¡Àº ´ÙÀ½ ÀڷḦ Âü°íÇϼ¼¿ä!
>>Gibson¡¯s method¿Í ¿ø¸®ºÎÅÍ °á°ú±îÁö ´Ù¸¥ In-Fusion¢ç Snap


* º» Á¦Ç°Àº In-Fusion¢ç HD Cloning kit (Code 639648)ÀÇ ¾÷±×·¹ÀÌµå ¹öÀü Á¦Ç°ÀÔ´Ï´Ù.

  • Subcloning ºÒÇÊ¿ä - Cloning À§Ä¡, insert ¼­¿­¿¡ °ü°è¾øÀÌ, ¸ñÀû vector¿¡ ¹Ù·Î cloning
  • ³ôÀº È¿À² - 0.5~15 kbÀÇ ´Ù¾çÇÑ Å©±âÀÇ fragment¿¡¼­ 95% ÀÌ»óÀÇ Á¤È®µµ
  • Seamless construction -ºÒÇÊ¿äÇÑ ¿°±â Ãß°¡ ¾øÀÌ cloning ¿Ï·á (e.g. Á¦ÇÑÈ¿¼Ò »ç¿ë ½Ã, TA cloning ½Ã)
  • ´Ù¾çÇÑ application - Single insert cloningÀº ¹°·Ð multiple cloning, mutagenesis µî Àû¿ë °¡´É (´õ ¸¹Àº Àû¿ë »ç·Ê¸¦ º¸½Ã·Á¸é Ŭ¸¯Çϼ¼¿ä)

Á¦Ç°¼³¸í
In-Fusion® Snap Assembly´Â ´Ü 15ºÐ ¹ÝÀÀÀ¸·Î ¼±ÇüÈ­ vector¿¡ ¾î¶² insert¶óµµ ºÒÇÊ¿äÇÑ ¿°±â Ãß°¡ ¾øÀÌ ¹æÇ⼺ ÀÖ´Â cloningÀÌ °¡´ÉÇÏ´Ù. Insert´Â Á¦ÇÑÈ¿¼Ò 󸮳ª phosphorylation, ȤÀº blunt-end ó¸® µî º°µµÀÇ ½ÇÇè °úÁ¤ÀÌ ÇÊ¿äÇÏÁö ¾Ê´Ù. In-Fusion® Snap AssemblyÀÇ È¿À²Àº 95% ÀÌ»óÀ¸·Î, ÃæºÐÇÑ ¼öÀÇ colony¸¦ ¾òÀ» ¼ö ÀÖ¾î ´ë·® ºÐ¼®À» À§ÇÑ scale up¿¡µµ Àû¿ëÇÒ ¼ö ÀÖ´Ù.
In-Fusion® Snap Assembly Master Mix´Â ligase ¾øÀ̵µ, insert¿Í ¼±ÇüÈ­ vector ¾ç ¸»´Ü 15 bpÀÇ »óµ¿ ¿°±â¼­¿­À» ÀÌ¿ëÇÏ¿© È¿À²ÀûÀ¸·Î Á¤È®ÇÏ°Ô °áÇÕ½ÃŲ´Ù. ¿Â¶óÀο¡¼­ Á¦°øÇÏ´Â In-Fusion® primer design toolÀ» ÀÌ¿ëÇϸé, °¢°¢ÀÇ ½ÇÇè¿¡ ¾Ë¸ÂÀº primer¸¦ ½±°Ô µðÀÚÀÎÇÒ ¼ö ÀÖ´Ù. ¹ÝÀÀÀÌ ½ÃÀ۵Ǹé, In-Fusion® Master Mix´Â ¼±ÇüÈ­µÈ DNA °¡´ÚÀÇ 3’ ¸»´ÜÀ¸·ÎºÎÅÍ ¿°±â¼­¿­À» Á¦°ÅÇÏ°í, µÎ °³ÀÇ DNA ´ÜÆí¿¡¼­ »óº¸¼­¿­ °£ÀÇ annealingÀ» ÅëÇØ DNA¸¦ °áÇÕ½ÃŲ´Ù. ÀÌ ¹æ¹ýÀº subcloningÀ̳ª º°µµÀÇ Á¶ÀÛ ¾øÀÌ, ¿©·¯ °³ÀÇ fragment¸¦ ÇϳªÀÇ vector¿¡ ¹Ù·Î Àû¿ëÇÒ ¼ö ÀÖ¾î multiple cloningÀÌ °¡´ÉÇÏ´Ù.
In-Fusion® Snap Assembly Master Mix¿¡´Â master mix¿Í control vector, control insert°¡ Æ÷ÇԵǾî ÀÖ¾î cloning È¿À²À» °°ÀÌ È®ÀÎÇÒ ¼ö Àֱ⠶§¹®¿¡ ½ÇÇèÀÇ ½Å·Ú¼º ¶ÇÇÑ È®ÀÎÀÌ °¡´ÉÇÏ´Ù.
Stellar™ Competent Cells (Code 636763 ¿Ü)Àº In-Fusion® ½Ã½ºÅÛ¿¡ ÃÖÀûÈ­µÇ¾î ÀÖÀ¸¸ç, In-Fusion® Snap Assembly Master Mix with Competent Cells (Code 638951, 638952)¿¡´Â Stellar™ Competent CellsÀÌ Æ÷ÇԵǾî ÀÖ´Ù.

±×¸² 1. In-Fusion® Workflow


±×¸² 2. In-Fusion® Snap Assembly¿Í In-Fusion HD (±âÁ¸ Á¦Ç°) °£ÀÇ È¿À² ºñ±³. In-Fusion® Snap Assembly´Â ±âÁ¸ Á¦Ç°¿¡ ºñÇØ Á¤È®µµ´Â À¯ÁöÇϸ鼭, colony Çü¼º ´É·ÂÀÌ °³¼±µÇ¾ú´Ù.
Á¦Ç° ¼º´É ºñ±³¸¦ À§ÇØ In-Fusion® HD (±âÁ¸ Á¦Ç°)°ú In-Fusion® Snap AssemblyÀ» »ç¿ëÇÏ¿© ¼­·Î ´Ù¸¥ Å©±âÀÇ inserts (405 bp-1,005 bp) 5°³¸¦ 2.7 kb vector¿¡ cloningÇÏ¿´´Ù. Cloning ¹ÝÀÀÀº °¢°¢ ¼¼ ¹ø ¹Ýº¹ ½ÇÇèÇÏ¿´°í, transformation ÈÄ °¢ ¹ÝÀÀÀ¸·ÎºÎÅÍ »ý¼ºµÈ colony¸¦ °¢ 20°³ ¼±ÅÃÇÏ¿© sanger sequencing ÇÑ °á°ú, In-Fusion® Snap Assembly¿¡¼­ ³ôÀº Á¤È®µµ¸¦ À¯ÁöÇϸ鼭 ±âÁ¸ Á¦Ç°º¸´Ù 4¹è ´õ ¸¹Àº colony¸¦ »ý¼ºÇÏ¿´´Ù. µÎ ½ÇÇè¿¡¼­ In-Fusion® master mix¸¦ Á¦¿ÜÇÑ primer¿Í ½Ã¾à µîÀº µ¿ÀÏÇÏ°Ô ÁøÇàÇÏ¿´´Ù.


±×¸² 3. In-Fusion® Snap Assembly¿Í NEBuilder HiFi (1). PCR·Î ¼±ÇüÈ­ÇÑ vector¿¡ single insert¸¦ cloningÇÑ °á°ú, insert Å©±â¿¡ °ü°è¾øÀÌ NEBuilder HiFiº¸´Ù In-Fusion® Snap Assembly¿¡¼­ ÈξÀ ¸¹Àº colony ¼ö¸¦ º¸¿´°í, 95% ÀÌ»óÀ¸·Î ³ôÀº Á¤È®µµ¸¦ º¸¿´´Ù.
3.8 kb insert (A), 34.2 kb adenovirus insert (B)¸¦ 2.7 kbÀÇ ¼±ÇüÈ­ º¤ÅÍ¿¡ cloningÇÏ¿´´Ù. Cloning ¹ÝÀÀÀº °¢°¢ ¼¼ ¹ø ¹Ýº¹ ½ÇÇèÇÏ¿´°í, transformation ÈÄ °¢ ¹ÝÀÀÀ¸·ÎºÎÅÍ »ý¼ºµÈ colony¸¦ 20°³ ¼±ÅÃÇÏ¿© Sanger sequencing°ú colony PCR ¹æ¹ýÀ¸·Î cloning Á¤È®µµ¸¦ È®ÀÎÇÏ¿´´Ù. ±× °á°ú, In-Fusion® Snap Assembly°¡ NEBuilder HiFi¿¡ ºñÇØ 2¹è ´õ ¸¹Àº colony¸¦ »ý¼ºÇÏ¿´´Ù.


±×¸² 4. In-Fusion® Snap Assembly¿Í NEBuilder HiFi (2). Á¦ÇÑÈ¿¼Ò·Î ¼±ÇüÈ­ÇÑ vector¿¡ single insert¸¦ cloningÇÑ °á°ú, ¸»´ÜÀÇ ÇüÅ (overhangs, blunt)¿¡ °ü°è¾øÀÌ NEBuilder HiFiº¸´Ù In-Fusion® Snap Assembly¿¡¼­ ÈξÀ ¸¹Àº colony ¼ö¸¦ º¸¿´´Ù.
In-Fusion® Snap Assembly¿Í Ÿ»ç Á¦Ç° (NEBuilder HiFi)¸¦ ÀÌ¿ëÇØ Á¦ÇÑÈ¿¼Ò¸¦ ó¸®ÇÏ¿© Á¦ÀÛÇÑ 5’ overhang (A), blunt end (B), 3’ overhang (C)ÀÇ ´ÜÀÏ insert (3.8 kb)¸¦ ¼±ÇüÈ­ÇÑ Ç¥ÁØ vector¿¡ cloningÇÏ¿´´Ù. Cloning ¹ÝÀÀÀº °¢°¢ ¼¼ ¹ø ¹Ýº¹ ½ÇÇèÇÏ¿´°í, °¢°¢ »ý¼ºµÈ colony¸¦ 20°³ ¼±ÅÃÇÏ¿© Sanger sequencingÀ¸·Î ¼­¿­ÀÌ Á¤È®ÇÏ°Ô »ðÀԵǾú´ÂÁö È®ÀÎÇÏ¿´´Ù.
Applications

±×¸² 5. In-Fusion® Snap Assembly Master MixÀÇ ´Ù¾çÇÑ applications ¿¹½Ã
ÀÚ¼¼ÇÑ ±¸¼ºÀº CoA¸¦ Âü°íÇϼ¼¿ä.
°ü·ÃÁ¦Ç°
- ÆíÀǼºÀÌ ÁÁÀº ½Ç¿Â º¸°ü master mix, In-Fusion® Snap Assembly EcoDry™ Master Mix (Code 638954)
- In-Fusion® °ú ¦²á, ÃÖ°íÀÇ Á¤È®µµ¸¦ Áö´Ñ PCRÈ¿¼Ò, PrimeSTAR® Max DNA Polymerase (Code R045A)
- In-Fusion® Cloning¿¡ ÃÖÀûÈ­µÈ °íÈ¿À² competent cells, Stellar™ Competent Cells (Code 636763)
- Touch down PCR ¹× gradient PCR ¼³Á¤ÀÌ °¡´ÉÇÑ °£ÆíÇÏ°í ÄÄÆÑÆ®ÇÑ PCR ±â±â, Clontech PCR Thermal Cycler GP (Code WN400)
- Plasmid DNA¸¦ ½Å¼ÓÇÏ°Ô Á¤Á¦ÇÒ ¼ö ÀÖ´Â, TaKaRa MiniBEST Plasmid Purification Kit (Code 9760A ¿Ü)

Keyword : infusion,in-fusion,cloning,PCR cloning,cloning,Ŭ·Î´×,º¹¼ö´ÜÆí,ÀÎÇ»Àü,ÇÁ·Î¹Í½º,premix, mutagenesis,mutation,soeasy,so easy,639648,639649,639650,636763,638909,638910,638911

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