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ÀüÁ¦Ç°º¸±â
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Cloning °ü·Ã
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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
Á¦Á¶»ç
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(ºÎ°¡¼¼º°µµ)
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Clontech
638947
In-Fusion¢ç Snap Assembly Master Mix
10 ȸ
270,000¿ø
,
Clontech
638948
In-Fusion¢ç Snap Assembly Master Mix
50 ȸ
1,150,000¿ø
Clontech
638949
In-Fusion¢ç Snap Assembly Master Mix
250 ȸ
3,739,000¿ø
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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
¢ç
Snap Assembly´Â 15ºÐ¸¸ÀÇ ¹ÝÀÀÀ¸·Î ¼±ÇüÈ vector¿¡ ¾î¶² insert¶óµµ ºÒÇÊ¿äÇÑ ¿°±â Ãß°¡ ¾øÀÌ ¹æÇ⼺ ÀÖ´Â cloningÀ» °¡´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
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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
¢ç
Snap Assembly Master MixÀº ligase ¾øÀ̵µ, insert¿Í ¼±ÇüÈ vector ¾ç ¸»´Ü 15 bpÀÇ »óµ¿ ¿°±â¼¿À» ÀÌ¿ëÇÏ¿© È¿À²ÀûÀ̰í Á¤È®ÇÏ°Ô °áÇÕ½ÃŲ´Ù. ¿Â¶óÀο¡¼ Á¦°øÇÏ´Â In-Fusion
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primer design toolÀ» ÀÌ¿ëÇϸé, °¢°¢ÀÇ ½ÇÇè¿¡ ¾Ë¸ÂÀº primer¸¦ ½±°Ô µðÀÚÀÎÇÒ ¼ö ÀÖ´Ù. ¹ÝÀÀÀÌ ½ÃÀ۵Ǹé, In-Fusion
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Master Mix´Â ¼±ÇüÈ µÈ DNA °¡´ÚÀÇ 3¡¯ end·ÎºÎÅÍ ¿°±â¼¿À» Á¦°ÅÇϰí, µÎ °³ÀÇ DNA ´ÜÆí¿¡¼ »óº¸¼¿ °£ÀÇ annealingÀ» ÅëÇØ DNA°¡ °áÇÕ½ÃŲ´Ù. ÀÌ ¹æ¹ýÀº SubcloningÀ̳ª º°µµÀÇ Á¶ÀÛ ¾øÀÌ, ÇϳªÀÇ fragment»Ó ¾Æ´Ï¶ó ¿©·¯ °³ÀÇ fragment¸¦ ÇϳªÀÇ vector¿¡ ¹Ù·Î Àû¿ëÇÒ ¼ö ÀÖ¾î high throughput application¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Ù.
±×¸² 1. 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¯ÁöÇϸé¼, colony Çü¼º ´É·ÂÀÌ °³¼±µÇ¾ú´Ù.
Five different inserts (ranging in size from 405 bp to 1,005 bp) were simultaneously cloned into a 2.7-kb vector previously linearized by inverse PCR using In-Fusion HD and In-Fusion Snap Assembly. Each cloning reaction was performed in triplicate. Amplification primers and all reagents, except for the cloning master mixes, were the same between the two systems. After transformation and plating, 10 colonies from each replicate were analyzed by Sanger sequencing to determine the cloning accuracy. In-Fusion Snap Assembly yielded 4X more colonies than In-Fusion HD while maintaining high cloning accuracy.
±×¸² 2. 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¸¿´´Ù. Á¤È®µµ´Â 95% ÀÌ»óÀ̾ú´Ù.
A single 3.8-kb insert (
Panel A
) or a 34.2-kb adenovirus insert (
Panel B
) was cloned into a 2.7-kb vector which was linearized via inverse PCR. These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers' specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing (for the 3.8-kb insert) or colony PCR (for the adenovirus insert) to determine the cloning accuracy. In-Fusion Snap Assembly yielded 2X more colonies than NEBuilder HiFi.
±×¸² 3. 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.
A single 3.8-kb insert was cloned into a standard cloning vector linearized by restriction digests to create 5' overhangs (
Panel A
), blunt ends (
Panel B
), or 3' overhangs (
Panel C
). These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers' specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing to determine the cloning accuracy. In-Fusion Snap Assembly yielded between 5X and 16X more colonies than NEBuilder HiFi, depending on the type of linearized vector ends.
Applications
- PCR cloning
- High-throughput cloning (see full range of reagent options on our high-throughput cloning page)
- Multiple-fragment cloning
- Gene synthesis
- Gene design
- Mutagenesis
- Domain swapping
- Domain modification
ÀÚ¼¼ÇÑ ±¸¼ºÀº CoA¸¦ Âü°íÇϼ¼¿ä.
Keyword : infusion,in-fusion,cloning,PCR cloning,cloning,Ŭ·Î´×,º¹¼ö´ÜÆí,ÀÎÇ»Àü,ÇÁ·Î¹Í½º,premix, mutagenesis,mutation,soeasy,so easy
In-Fusion Cloning
[Overview] In-Fusion¢ç HD Cloning
[Àû¿ë] In-Fusion¢ç HD Cloning Àû¿ë»ç·Ê
Online Tools for In-Fusion
[FAQ] In-Fusion¢ç HD Cloning
½ÇÁ¦ °í°´ÀÇ »ç¿ëÈıâ
2ź ½ÇÁ¦ »ç¿ëÀÚ Èıâ
[°³¿ä] So Easy Product
ÃÖ°íÀÇ Colony Çü¼º´É°ú Á¤È®ÇÑ PCR cloning
[NEW] In-Fusion¢ç Snap Assembly
PremixÈ! ½Ã°£´ÜÃà! È¿À²¼º Çâ»ó!
[Kit] In-Fusion¢ç Cloning
[Kit] In-Fusion¢ç EcoDry Cloning
[Plus] In-Fusion¢ç Cloning
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In-Fusion¢ç Ready Vectors