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 Á¦¸ñ : [À̺¥Æ® Á¾·á] [ÇÁ·Î¸ð¼Ç] Ÿ»ç´ëºñ °¡Àå Á¤È®ÇÏ°í ºü¸¥ In-Fusion cloning 20% ÇÒÀÎ
ÀÛ¼ºÀÏ : 2024.06.07
 ÀÛ¼ºÀÚ : ´ÙÄ«¶óÄÚ¸®¾Æ

¾î¶² ¹æ¹ýº¸´Ù ½±°í ºü¸£´Ù°¡Àå Á¤È®ÇÑ cloning ¹æ¹ý Ãßõ, 

In-Fusion¢ç Snap Assembly Master Mix 20% ÇÒÀÎ ±âȸ!

 

±â°£: 2024³â 5¿ù 2ÀÏ (¸ñ) ~ 6¿ù 28ÀÏ (±Ý)

 

¡á In-Fusion¢ç ±â¼ú ¼Ò°³

3¡¯ ¡æ 5¡¯ exonuclease È°¼ºÀ» °¡Áö´Â In-Fusion¢ç È¿¼Ò¸¦ ÀÌ¿ëÇØ DNA ´ÜÆí °£ÀÇ »óµ¿¼­¿­ (¾à 15 bp)¸¦ À¶ÇÕ½ÃÄÑ cloningÇϴ ±â¼ú·ÎÁ¦ÇÑÈ¿¼Ò/ligase ¾øÀ̵µ ½±°í Á¤È®ÇÏ°Ô seamlessÇÑ »ê¹°À» ¾òÀ» ¼ö ÀÖ´Ù.

 

>> In-Fusion¢ç ¿ø¸® µ¿¿µ»ó 

>> In-Fusion¢ç primer µðÀÚÀΠÅø 

>> In-Fusion¢ç Cloning Àû¿ë»ç·Ê

 

¡á In-Fusion¢ç Å¸»ç´ëºñ ÀåÁ¡

¡æ In-Fusion¢ç Snap Assembly¿Í Gibson assembly ºñ±³ 

 

In-Fusion¢ç Snap Assembly

Gibson assembly-based cloning

Colony ¼ö

ÀϹÝÀûÀΠcloningºÎÅÍ *º¹ÀâÇÑ cloning±îÁö Ÿ ¹æ½Ä¿¡ ºñÇØ colony°¡ ¸¹ÀÌ Çü¼ºµÊ

*º¹ÀâÇÑ cloningÀÇ °æ¿ì colony°¡ Àû°Ô Çü¼ºµÇ°Å³ªÇü¼ºµÇÁö ¾ÊÀ½

Nick ¼öº¹ °úÁ¤

DNA polymerase¸¦ »ç¿ëÇÏÁö ¾Ê°í, comp. cell¿¡¼­ nickÀ» ¼öº¹ÇÏ¿©, error°¡ °ÅÀÇ ¾øÀ½

DNA polymerase»ç¿ëÀ¸·Î nick ¼öº¹ Áß error°¡ ¹ß»ýµÉ ¼ö ÀÖÀ½

Ligase¸¦ »ç¿ëÇÏÁö ¾Ê¾Æ, self-ligation µîÀÇ ¹é±×¶ó¿îµå°¡ °ÅÀÇ ¾øÀ½

in vitro ligationÀ» ÀÌ¿ëÇϹǷÎ, self-ligationÀÇ È®·üÀÌ ³ôÀ½

¹ÝÀÀ ½Ã°£

15ºÐ

~60ºÐ

A-overhang insert ±³Á¤

3¡¯ ¡æ 5¡¯ exonuclease È°¼º:

PCR °úÁ¤¿¡¼­ ÀÓÀÇ·Î ºÎ°¡µÈ A 1¿°±âÀÇ ±³Á¤ °¡´É

5¡¯ ¡æ 3¡¯ exonuclease È°¼º:

PCR °úÁ¤¿¡¼­ ÀÓÀÇ·Î ºÎ°¡µÈ A 1¿°±âÀÇ ±³Á¤ ºÒ°¡´É (Á¤È®µµ°¡ ¶³¾îÁü)

PCR primer µðÀÚÀÎ

15 bp overlapÀ¸·Î, PCR Á¶°Ç ¼³Á¤ ½Ã À¯¸®

ÀϹÝÀûÀ¸·Î 20 ~ 30 bp overlap

*º¹ÀâÇÑ cloning: multiple cloning, large insert/vector µî

 

¡æ In-Fusion¢ç Snap Assembly¿Í NEBuilder HiFi ºñ±³ µ¥ÀÌÅÍ


±×¸²1. Insert Å©±â¿¡ °ü°è¾øÀÌ NEBuilder HiFiº¸´Ù In-Fusion¢ç Snap Assembly¿¡¼­ ÈξÀ ¸¹Àº colony ¼ö¸¦ º¸¿´°í, 95% ÀÌ»óÀ¸·Î ³ôÀº Á¤È®µµ¸¦ º¸¿´´Ù.



±×¸²2. Á¦ÇÑÈ¿¼Ò·Î ¼±ÇüÈ­ÇÑ vector¿¡ single insert¸¦ cloningÇÑ °á°ú¸»´ÜÀÇ ÇüÅ (overhangs, blunt)¿¡ °ü°è¾øÀÌ NEBuilder HiFiº¸´Ù In-Fusion¢ç Snap Assembly¿¡¼­ ÈξÀ ¸¹Àº colony ¼ö¸¦ º¸¿´´Ù.


¡á In-Fusion¢ç Á¦Ç° ¶óÀÎ

Code

Á¦Ç°¸í

Àû¿ë

638947

In-Fusion¢ç Snap Assembly Master Mix

 ¿øÇÏ´Â ´ë·Î ºü¸£°í °£´ÜÇÏ°Ô In-Fusion¢ç Cloning

638954

In-Fusion¢ç Snap Assembly EcoDry¢â Master Mix

 µ¿°á°ÇÁ¶ ÇüÅ·Π½Ç¿Âº¸°üÀÌ °¡´É

 ºü¸£°í °£´ÜÇÏ°Ô In-Fusion¢ç Cloning

638951

In-Fusion¢ç Snap Assembly Master Mix with Competent Cells

 ÃÖÀûÈ­µÈ comp. cellÀÌ Æ÷ÇÔµÈ ¹öÀüÀÇ In-Fusion¢ç Cloning

636763

Stellar¢â Competent Cells

 ³ôÀº ÇüÁúÀüȯ È¿À²À» °¡Áø E. coli competent cell

 

°ü·Ã ¹®ÀÇ»çÇ×Àº ´ÙÄ«¶óÄÚ¸®¾Æ Áö¿ª ´ë¸®Á¡ (´ë¸®Á¡ ¾È³»¶Ç´Â °í°´Áö¿ø (02-2081-2510)À¸·Î ¹®ÀÇ ¹Ù¶ø´Ï´Ù.

°¨»çÇÕ´Ï´Ù.

  


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