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Home > ÀüÁ¦Ç°º¸±â > Cloning °ü·Ã > Competent Cells & Electro-Cells > E. coli Competent Cells (Takara Bio Á¦Ç°)
Long strand DNAµµ methylated DNAµµ OK£¡°íÈ¿À² Competent Cell

E. coli Competent Cells (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
9051
E. coli HB101 Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(100 ¥ìl¡¿10)
180,800¿ø 
226,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9051_e.v1807Da.pdf
Takara
9052
E. coli JM109 Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(100 ¥ìl¡¿10)
180,800¿ø 
226,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9052_e.v2001Da.pdf
Takara
9053
E. coli CJ236 Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(100 ¥ìl¡¿10)
198,400¿ø 
248,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9053_e.v2012Da.pdf
Takara
9057
E. coli DH5¥á Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(100 ¥ìl¡¿10)
180,800¿ø 
226,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9057_e.v2001Da.pdf
Takara
9128
E. coli HST08 Premium Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(100 ¥ìl¡¿10)
208,000¿ø 
260,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9128_e.v2007Da.pdf
Takara
9129
E. coli HST04 dam£­/dcm£­ Competent Cells
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(100 ¥ìl¡¿10)
205,600¿ø 
257,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
9129_e.v0906Da.pdf

Á¦Ç°¼³¸í
Mandel°ú Higa¿¡ ÀÇÇØ È®¸³µÈ E. coliÀÇ ÇüÁúÀüȯ¹ýÀº ÀçÁ¶ÇÕ DNA ½ÇÇèÀÇ Áß¿äÇÑ ±âÃÊ°¡ µÇ¾ú´Ù. E. coli¸¦ Ca2+ ÀÌ¿Â Á¸Àç ÇÏ¿¡ ³õ¾ÆµÎ¸é ¿Ü·¡ DNA (plasmid, phage DNA µî)¸¦ ¼¼Æ÷ ¾ÈÀ¸·Î µµÀÔÇÒ ¼ö ÀÖ´Â "competent"»óÅ°¡ µÇ°í ÀÌ ¼¼Æ÷¸¦ competent cellÀ̶ó ÇÑ´Ù.
ÀçÁ¶ÇÕ DNA ½ÇÇè¿¡¼­´Â competent cellÀ» »ç¿ëÇÑ ÇüÁúÀüȯ, ÇüÁúµµÀÔ µîÀ» ¹ø¹øÇÏ°Ô ½Ç½ÃÇϳª, È¿À²ÀÌ ³ô°í ÀçÇö¼ºÀÌ ÁÁÀº competent cellÀ» Á¦Á¶ÇÏ´Â °ÍÀº ½±Áö ¾Ê´Ù. À¯ÀüÀÚ library Á¦ÀÛ, ÀçÁ¶ÇÕ plasmid Á¦ÀÛÀ̳ª subcloning µîÀ» ÇÒ ¶§, À¯ÀüÀÚ¾çÀÌ ÀûÀº °æ¿ì¿¡´Â µµÀÔ È¿À²ÀÌ ³ôÀº competent cellÀ» »ç¿ëÇÏ´Â °ÍÀÌ Áß¿äÇÏ´Ù.
´ÙÄ«¶ó¿¡¼­´Â HanahanÀÇ ¹æ¹ýÀ» °³·®ÇÑ »õ·Î¿î ¹æ¹ýÀ¸·Î ÇüÁúÀüȯ È¿À²ÀÌ ³ôÀº 7Á¾·ùÀÇ E. coli Competent Cells HST08 Premium, JM109, DH5¥á, HB101, HST04 dam£­/dcm£­, MV1184, CJ236 (¸ðµÎ EK1°èÀÇ ¼÷ÁÖ E. coli)¸¦ Á¦Á¶¡¤ ÆǸÅÇÏ°í ÀÖ´Ù.
±ÕÁÖ Æ¯Â¡
°íÈ¿À² ¥á-»óº¸¼º ¼±Åà ¼÷ÁÖ E. coli HST08 Premium Competent Cells (Code 9128)
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 cloning ºÎÅÍ À¯ÀüÀÚ libraryÀÇ Á¦ÀÛ, subcloningµî¿¡ 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 kb ÀÌ»óÀÇ DNA insertÀÇ cloning, library Á¦ÀÛ µîÀ» 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±°í, F-À̱⠶§¹®¿¡, BAC, fosmid vetctor¿¡µµ »ç¿ë °¡´ÉÇÏ´Ù.

¥á- »óº¸¼º ¼±Åà ¼÷ÁÖ E. coli JM109 Competent Cells (Code 9052)
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 Competent Cells (Code 9057)
pUC°è plasmid¸¦ ÀÌ¿ëÇÑ À¯ÀüÀÚ library Á¦ÀÛ, subcloningµîÀ» 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 Competent Cells (Code 9051)
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.

dam, dcm °á¼Õ ¼÷ÁÖ E. coli HST04 dam-/dcm- Competent Cells (Code 9129)
E. coli HST04 dam-/dcm-´Â º»·¡ E. coli°¡ °¡Áö°í ÀÖ´Â ¸ÞÆ¿È­ À¯ÀüÀÚ dam, dcmÀÌ °á¼ÕµÇ¾î ÀÖ¾î, DNA°¡ E. coli¿¡ ÀÇÇØ ¸ÞÆ¿È­µÇÁö ¾Ê´Â´Ù. º» Á¦Ç°À¸·Î Á¶Á¦µÈ plasmid´Â dam ¶Ç´Â dcm methylaseÀÇ ¿µÇâÀ» ¹Þ´Â Á¦ÇÑÈ¿¼Ò·Î Àý´Ü °¡´ÉÇÏ´Ù. dam, recAÀÌ °°ÀÌ °á¼Õ µÇ´Â °æ¿ì ¼¼Æ÷°¡ »ç¸êÇϱ⠶§¹®¿¡ ÀÌ ¼¼Æ÷´Â recA À¯ÀüÀÚ¸¦ ¹ßÇöÇÏ°í ÀÖ´Ù. ¹Ýº¹¼­¿­À» °®°í ÀÖ´Â ¿Ü·¡ DNA´Â recA À¯ÀüÀÚ¿¡ ÀÇÇØ ÀçÁ¶ÇÕÀÌ ÀϾ °¡´É¼ºÀÌ ÀÖÀ¸¹Ç·Î cloning¿ë ¼÷Áַδ ÀûÇÕÇÏÁö ¾ÊÀ¸¸ç, ÇüÁúÀüȯ½Ã¿¡´Â ¹Ì¸® ±¸ÃàµÈ plasmid¸¦ ÃßõÇÑ´Ù.

Amber º¯ÀÌ ¼±Åÿë ÇÑ°¡´Ú ssDNA Á¦Á¶¿ë ¼÷ÁÖ E. coli MV1184 Competent Cells (Code 9055)
E. coli MV1184´Â amber suppressor free ±ÕÁÖ·Î amber º¯ÀÌ DNAÀÇ ¼±º°¿¡ ÀÌ¿ëÇÒ ¼ö ÀÖ´Ù. ¶ÇÇÑ F¡¯ plasmid¸¦ °¡Áö±â ¶§¹®¿¡ M13 phage vector³ª phagemid vectorÀÇ ¼÷ÁÖ·Î ssDNAÀÇ Á¶Á¦¿¡µµ ÀÌ¿ëÇÒ ¼ö ÀÖ´Ù.

ºÎÀ§ ƯÀÌÀû º¯ÀÌ Ã³¸®¿ë ssDNA Á¦Á¶ ¼÷ÁÖ E. coli CJ236 Competent Cells (Code 9053)
E. coli CJ236Àº dut, ung º¯ÀÌÁÖ¸¦ °¡Áø E. coli¿¡¼­ dUTPase (Dut)¿Í Uracil-DNA glycosylase (Ung)°¡ °á¼ÕµÇ¾î ÀÖ¾î DNA Áß Thymine (T)ÀÇ ÀϺΰ¡ deoxyuracil (dU)·Î ġȯµÈ DNA¸¦ ÇÕ¼ºÇÑ´Ù. Kunkel¹ýÀº ÀÌ·¯ÇÑ E. coli¸¦ ÀÌ¿ëÇÏ¿© site-directed mutagenesis¸¦ ½Ç½ÃÇÏ´Â ¹æ¹ýÀ¸·Î, º¯ÀÌ ½ÃÅ°°íÀÚ ÇÏ´Â À¯ÀüÀÚ¸¦ pUC118/119, M13 phage µîÀÇ ssDNA¸¦ Á¶Á¦ÇÒ ¼ö ÀÖ´Â vector¿¡ cloningÇÏ¿© CJ236À» ¼÷ÁÖ·Î ssDNA¸¦ Á¶Á¦Çϸé Kunkel¹ýÀ¸·Î »ç¿ëÇÒ ¼ö ÀÖ´Â deoxyuracil ÇÔÀ¯ DNA¸¦ ¾òÀ» ¼ö ÀÖ´Ù. F¡Ç plasmid (pCJ105)¿¡ chloramphenicol ³»¼º À¯ÀüÀÚ¸¦ °®°í Àֱ⠶§¹®¿¡ chloramphenicol Á¸Àç ÇÏ¿¡¼­ÀÇ ¹è¾çÀÌ ÇÊ¿äÇÏ´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

E. coli HST04 dam-/dcm-

F-, ara¥Ä(lac-proAB)[¥Õ80dlacZ¥ÄM15], rpsL (str), thi, ¥Ä(mrr-hsdRMSmcrBC), ¥ÄmcrA, dam, dcm

E. coli MV1184

ara¥Ä(lac-proAB), rpsLthi (¥Õ80 lacZ¥ÄM15), ¥Ä(srl-recA)306:Tn10 (tetr)/F¡¯¡²traD36, proAB+lac Iq, lacZ¥ÄM15¡³

E. coli CJ236

dut1, ung1, thi-1, relA1/pCJ105 (F¡¯ camr)

ÇüÁúÀüȯ È¿À²
1 ng plasmid DNA¸¦ 100 §¡ÀÇ ¼¼Æ÷¿¡ transformationÇÑ °æ¿ì
HST08 Premium Competent Cells

>1 ¡¿ 108 cfu/§¶ pUC19

JM109 Competent Cells

>1 ¡¿ 108 cfu/§¶ pBR322

DH5¥á Competent Cells

>1 ¡¿ 108 cfu/§¶ pUC19

HB101 Competent Cells

>1 ¡¿ 108 cfu/§¶ pBR322

HST04 dam-/dcm- Competent Cells

>1 ¡¿ 106 cfu/§¶ pUC19

MV1184 Competent Cells

>1 ¡¿ 107 cfu/§¶ pUC119

CJ236 Competent Cells

>1 ¡¿ 107 cfu/§¶ pUC119

F¡¯-plasmidÀÇ ¾ÈÁ¤¼º, ¥á »óº¸¼º È®ÀÎ
- E. coli JM109, E.coli MV1184
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.
- E. coli CJ236
30 §¶/§¢ÀÇ chloramphenicolÀ» Æ÷ÇÔÇÑ ¹èÁö¿¡¼­ F¡¯-plasmid (pCJ105)°¡ ÇüÁúÀüȯ µÇÁö ¾ÊÀº E. coli´Â »ýÁ¸ÇÒ ¼ö ¾øÀ½À» È®ÀÎÇÏ¿´´Ù.
Cell ³óµµ 1~2 x 109 bacteria/§¢
Plasmid ½ºÅ©¸®´×¿¡ »ç¿ëµÇ´Â Ç×»ýÁ¦¿Í ³óµµ (§¶/§¢)

Code

Á¦Ç°¸í

Ampicillin

Kanamycin

Chloram-phenicol

Strepto-mycin

Tetracycline

9021

E. coli HB101 Electro-Cells

100

50

20

NA

20

9022

E. coli JM109 Electro-Cells

100

50

20

50

20

9027

E. coli DH5¥á Electro-Cells

100

50

20

50

20

9028

E. coli HST08 Premium Electro-Cells

100

50

20

50

20

9051

E. coli HB101 Competent Cells

100

50

20

NA

20

9052

E. coli JM109 Competent Cells

100

50

20

50

20

9053

E. coli CJ236 Competent Cells

100

50

NA

50

20

9055

E. coli MV1184 Competent Cells

100

50

20

NA

NA

9057

E. coli DH5¥á Competent Cells

100

50

20

50

20

9128

E. coli HST08 Premium Competent Cells

100

50

20

50

20

9129

E. coli HST04 dam£­/ dcm£­ 
Competent Cells

100

50

20

NA

20

°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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