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Home > ÀüÁ¦Ç°º¸±â > Cloning °ü·Ã > Competent Cells & Electro-Cells > E. coli Electro-Cells À϶÷ (Takara Bio Á¦Ç°)

E. coli Electro-Cells À϶÷ (Takara Bio Á¦Ç°)

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
9021
E. coli HB101 Electro-Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(50 ¥ìl¡¿10)
313,000¿ø  9021_e.v2010Da.pdf
Takara
9022
E. coli JM109 Electro-Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(50 ¥ìl¡¿10)
313,000¿ø  9022_e.v2101Da.pdf
Takara
9027
E. coli DH5¥á Electro-Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(50 ¥ìl¡¿10)
260,000¿ø  9027_e.v2002Da.pdf
Takara
9028
E. coli HST08 Premium Electro-Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(50 ¥ìl¡¿10)
290,000¿ø  9028_e.v2012Da.pdf

Á¦Ç°¼³¸í
°íÀü¾Ð pulse·Î ¼¼Æ÷¸·¿¡ ÀϽÃÀûÀ̸鼭 °¡¿ªÀûÀ¸·Î ±Õ¿­À» ÀÏÀ¸ÄÑ DNA¸¦ ¼¼Æ÷³»·Î À̵¿½ÃÅ°´Â electroporation¹ýÀº ¿©·¯ °¡Áö ÇüÁúÀüȯ¹ý Áß °¡Àå È¿°úÀûÀÎ ¹æ¹ý Áß ÇϳªÀÌ´Ù. Á¾·¡ÀÇ Ca2+ Á¸Àç ÇÏ¿¡¼­ Á¶Á¦ÇÑ competent cell¿¡ ºñÇØ ÇüÁúÀüȯ È¿À²°ú °á°úÀÇ ÀçÇö¼ºÀÌ ¿ì¼öÇÏ¿© ½ÇÇè½Ã°£À» Àý¾àÇÒ »Ó¸¸ ¾Æ´Ï¶ó ¼Ò·®ÀÇ ½Ã·á¸¦ ´ëÀå±Õ¿¡ µµÀÔÇÒ ¶§ ƯÈ÷ À¯¿ëÇÏ´Ù.
´ÙÄ«¶ó¿¡¼­´Â µ¶ÀÚÀûÀÎ ¹è¾ç¹ýÀ¸·Î ÇüÁúÀüȯ È¿À²ÀÌ º¸´Ù ³ôÀº 4Á¾·ùÀÇ Electro-Cells E. coli HST08 Premium, JM109, DH5¥á, HB101¸¦ Á¦Á¶, ÆǸÅÇÏ°í ÀÖ´Ù.

(ÁÖÀÇ) º» Á¦Ç°Àº Heat shock¿¡ ÀÇÇÑ ÇüÁúÀüȯ¿¡´Â »ç¿ëÇÒ ¼ö ¾ø´Ù. ÀÌ °æ¿ì E. coli Competent Cells (Code 9128¿Ü)ÀÇ Á¦Ç°À» »ç¿ëÇÑ´Ù.
±ÕÁÖ Æ¯Â¡
°íÈ¿À² ¥á-»óº¸¼º ¼±Åà ¼÷ÁÖ E. coli HST08 Premium Electro-Cells (Code 9028)
E. coli HST08 Premium´Â ¿Ü·¡ÀÇ ¸ÞÆ¿È­µÈ DNA¸¦ Àý´ÜÇÏ´Â À¯ÀüÀÚ±º mrr, hsdRMS, mcrBC, mcrAÀÌ °á¼ÕµÇ¾î ÀÖ´Ù. ¸Å¿ì ³ôÀº ÇüÁú Àüȯ ´É·ÂÀ» °¡Áö°í Àֱ⠶§¹®¿¡ ¸ÞÆ¿È­µÈ DNAÀÇ Å¬·Î´×ºÎÅÍ À¯ÀüÀÚ ¶óÀ̺귯¸®ÀÇ Á¦ÀÛ, ¼­ºê Ŭ·Î´× µî¿¡ À̸£±â±îÁö Æø³ÐÀº ¿ëµµ¿¡ »ç¿ëÇÒ ¼ö ÀÖ´Ù. Å« »çÀÌÁîÀÇ plasmid DNAÀÇ ÇüÁú Àüȯµµ ³ôÀº È¿À²·Î °¡´ÉÇÏ°í, °°Àº À¯ÀüÀÚÇüÀ» °¡Áö´Â ´Ù¸¥ competent cellsº¸´Ù colony Çü¼º ¼Óµµµµ ºü¸£±â ¶§¹®¿¡, TaKaRa DNA Ligation Kit LONG(Code 6024)°ú °°ÀÌ »ç¿ëÇϸé, 10 kbÀÌ»óÀÇ DNAÀÇ Å¬·Î´×, ¶óÀ̺귯¸® Á¦ÀÛµîÀ» º¸´Ù ½±°Ô ÇÒ ¼ö ÀÖ´Ù. pUC°è plasmid¸¦ ÀÌ¿ëÇÑ ÇüÁú Àüȯ ½Ã¿¡´Â, ¥â-galatosidaseÀÇ ¥á-»óº¸¼ºÀ» ÀÌ¿ëÇØ, X-Gal ÷°¡¿¡ ÀÇÇÑ ÀçÁ¶ÇÕüÀÇ ¼±º°ÀÌ ½±´Ù. F-À̱⠶§¹®¿¡, BAC, fosmid vetctor¿¡µµ »ç¿ë °¡´ÉÇÏ´Ù.

¥á- »óº¸¼º ¼±Åà ¼÷ÁÖ E. coli JM109 Electro-Cells (Code9022)
E. coli JM109´Â pUC°è plasmid vectorÀÇ ÇüÁúÀüȯÀ̳ª M13 phage vector DNAÀÇ ÇüÁúµµÀÔ µîÀ» ÇÏ´Â °æ¿ì, vector·ÎºÎÅÍ ¹ßÇöÇÏ´Â lacZ ¥á peptide¿Í JM109F¡Ç°¡ ÄÚµùÇÏ´Â lacZ¡âM15¿¡ ÀÇÇÑ ¥â-galactosidaseÀÇ È°¼ºÈ¸º¹ (¥á-»óº¸¼º)À» ÀÌ¿ëÇÔÀ¸·Î½á ÀçÁ¶ÇÕüÀÇ ¼±º°ÀÌ °£ÆíÇÑ ±ÕÁÖÀÌ´Ù. F¡Ç plasmid¸¦ °®°í Àֱ⠶§¹®¿¡ À¯ÀüÀÚ library Á¦ÀÛÀ̳ª subcloning ÀÌ¿Ü¿¡ M13 vector DNAÀÇ ¼÷Áַνá ssDNAÀÇ Á¦Á¶¿¡µµ »ç¿ëÇÒ ¼ö ÀÖ´Ù.

¥á- »óº¸¼º ¼±Åà ¼÷ÁÖ E. coli DH5¥á Electro-Cells (Code 9027)
pUC°è plasmid¸¦ ÀÌ¿ëÇÑ À¯ÀüÀÚ library Á¦ÀÛ, ¼­ºê Ŭ·Î´× µîÀ» ÇÏ´Â °æ¿ì, º» ¼÷ÁÖ±Õ°ú Á¶ÇÕÇÏ¸é ¥â-galactosidaseÀÇ ¥á-»óº¸¼ºÀ» ÀÌ¿ëÇÏ¿© X-Gal¿¡ ÀÇÇØ ÀçÁ¶ÇÕü¸¦ ½±°Ô ¼±º°ÇÒ ¼ö ÀÖ´Ù. deoR º¯ÀÌ°¡ Æ÷ÇԵǾî Àֱ⠶§¹®¿¡ Å« Å©±âÀÇ PlasmidÀÇ µµÀÔÀÌ °¡´ÉÇÏ´Ù.

°íÈ¿À² ¹ü¿ë ¼÷ÁÖ E. coli HB101 Electro-cells (Code 9021)
E. coli HB101Àº ÀçÁ¶ÇÕ DNA ½ÇÇèÀÇ ÃʱâºÎÅÍ ³Î¸® »ç¿ë µÇ¾î¿Â ±ÕÁÖ·Î À¯ÀüÇüÁúÀÌ ¾ÈÁ¤ÀûÀÌ°í »ç¿ëÀÌ ¿ëÀÌÇÏ¿© ÇüÁúÀüȯ ½ÇÇè ¸ñÀû¿¡ Àû´çÇÏ´Ù.
º¸Á¸ -80¡É
Genotype

Strain

Genotype

E. coli HST08 Premium

F-, endA1, supE44, thi-1, recA1, relA1, gyrA96, phoA, ¥Õ80dlacZ¥ÄM15, ¥Ä(lacZYA-argF)U169, ¥Ä(mrr-hsdRMS-mcrBC), ¥ÄmcrA, ¥ë-

E. coli JM109

recA1, endA1, gyrA96, thi-1, hsdR17 (rK-, mK-), e14- (mcrA-), supE44, relA1, ¥Ä(lac-proAB)/F¡¯ ¡²traD36, proAB+, lacIq, lacZ¥ÄM15¡³

E. coli DH5¥á

F-, ¥Õ80dlacZ¥ÄM15, ¥Ä(lacZYA-argF) U169, deoRrecA1, endA1, hsdR17 (rK-, mK+), phoAsupE44, ¥ë-, thi-1, gyrA96, relA1

E. coli HB101

supE44, hsdS20(rB-, mB-), recA13, ara-14, proA2, lacY1, galK2, rpsL20, xyl-5, mtl-1, leuB6, thi-1

ÇüÁúÀüȯ È¿À²
10 pg plasmid DNA¸¦ 50 §¡ÀÇ ¼¼Æ÷¿¡ transformationÇÑ °æ¿ì
HST08 Premium Competent Cells

>1 ¡¿ 109 cfu/§¶ pUC19

JM109 Competent Cells

>1 ¡¿ 109 cfu/§¶ pUC19

DH5¥á Competent Cells

>1 ¡¿ 109 cfu/§¶ pUC19

HB101 Competent Cells

>1 ¡¿ 108 cfu/§¶ pBR322

F¡¯ plasmidÀÇ ¾ÈÁ¤¼º, ¥á »óº¸¼º È®ÀÎ
- E. coli JM109
pUC19À» ÀÌ¿ëÇØ ÇüÁúÀüȯÀ» ÇÑ ÈÄ 100 §¶/§¢ ampicillin, 0.3 mM IPTG, 60 §¶/§¢ X-GalÀ» Æ÷ÇÔÇÑ LB-agar ¹èÁö¿¡ µµ¸»ÇÑ °æ¿ì white colony´Â 10% ÀÌÇÏ·Î ¹ß»ýÇÏ¿´´Ù.
- E. coli HST08 Premium, E.coli DH5¥á
pUC19À» ÀÌ¿ëÇØ ÇüÁúÀüȯÀ» ÇÑ ÈÄ 100 §¶/§¢ ampicillin, 60 §¶/§¢ X-GalÀ» Æ÷ÇÔÇÑ LB-agar ¹èÁö¿¡ µµ¸»ÇÑ °æ¿ì blue colony¸¦ È®ÀÎÇÒ ¼ö ÀÖ´Ù.
Cell ³óµµ > x 1010 bacteria/§¢
¼øµµ
50 §¡ÀÇ E. coli Electro-CellsÀ» 100 §¶/§¢ÀÇ ampicillinÀÌ Æ÷ÇÔµÈ LB-agar ¹èÁö¿¡ Á¢Á¾ÇÑ °æ¿ì colony°¡ Çü¼ºµÇÁö ¾Ê´Â´Ù.
°ü·ÃÁ¦Ç°
- Cloning¿¡ µü¸Â´Â ¹ü¿ë¼º ³ôÀº high fidelity PCR È¿¼Ò PrimeSTAR¢ç GXL DNA Polymerase (Code R050A)
- °íÈ¿À² ligation premix DNA Ligation Kit £¼Mighty Mix£¾(Code 6023)
- TA cloning ¿ë T-vector¿Í ligation premix Mighty TA-cloning Kit (Code 6028)
- ¹ÚÅ׸®¾Æ¿¡¼­ plasmid DNA Á¤Á¦ TaKaRa MiniBEST Plasmid Purification Kit (9760A)

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