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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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Takara
9051
E. coli HB101 Competent Cells
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1 Set
(100 ¥ìl¡¿10)
207,000¿ø  44806
Takara
9052
E. coli JM109 Competent Cells
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1 Set
(100 ¥ìl¡¿10)
207,000¿ø  43114
Takara
9053
E. coli CJ236 Competent Cells
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1 Set
(100 ¥ìl¡¿10)
227,000¿ø  44826
Takara
9054
E. coli BMH71-18 mutS Competent Cells
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1 Set
(100 ¥ìl¡¿10)
227,000¿ø  10319
Takara
9055
E. coli MV1184 Competent Cells
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1 Set
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227,000¿ø  44828
Takara
9057
E. coli DH5¥á Competent Cells
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1 Set
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207,000¿ø  44846
Takara
9127
E. coli HST02 Competent Cells
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1 Set
(100 ¥ìl¡¿10)
283,000¿ø  10337
Takara
9128
E. coli HST08 Premium Competent Cells
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1 Set
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239,000¿ø  64501
Takara
9129
E. coli HST04 dam£­/dcm£­ Competent Cells
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1 Set
(100 ¥ìl¡¿10)
236,000¿ø  9129_e.v0906Da.pdf

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Á¦Ç°¼³¸í
Mandel°ú Higa¿¡ ÀÇÇÑ ´ëÀå±ÕÀÇ ÇüÁúÀüȯ¹ýÀÇ È®¸³Àº ÀçÁ¶ÇÕ DNA ½ÇÇèÀÇ Áß¿äÇÑ ±âÃʰ¡ µÇ¾ú´Ù. ´ëÀå±ÕÀ» Ca2+ À̿ Á¸Àç ÇÏ¿¡ ³õ¾ÆµÎ¸é ¿Ü·¡ DNA (plasmid, phage DNA µî)¸¦ ¼¼Æ÷ ¾ÈÀ¸·Î µµÀÔÇÒ ¼ö ÀÖ´Â "competent"»óŰ¡ 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 ¼­ºêŬ·Î´× µî ¸ñÀûÀ¸·Î ÇÏ´Â 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 10Á¾·ùÀÇ Competent Cells E. coli HST08 Premium, HST04 dam£­/dcm£­, HB101, JM109, DH5¥á, CJ236, BMH71-18 mutS, MV1184, TH2, HST02 (¸ðµÎ EK1°èÀÇ ¼÷ÁÖ ´ëÀå±ÕÀÓ)¸¦ Á¦Á¶¡¤ ÆÇ¸ÅÇϰí ÀÖ´Ù.

*¸Þƿȭ DNAÀÇ Å¬·Î´×, cDNA library, genome DNA library Á¦ÀÛºÎÅÍ ÀϹÝÀûÀÎ cloning±îÁö Æø³Ð°Ô Àû¿ëÇÏ´Â E. coli HST08 Premium Competent Cells (Code 9128) ÀÚ¼¼È÷ º¸±â
±ÕÁÖ Æ¯Â¡
°íÈ¿À² ¥á-»óº¸¼º ¼±Åà ¼÷ÁÖ E. coli HST08 Premium
E. coli HST08 Premium´Â ¿Ü·¡ÀÇ ¸ÞƿȭµÈ 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®¿¡ ¸ÞƿȭµÈ 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 °¡´ÉÇϰí, °°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.

dam, dcm °á¼Õ ¼÷ÁÖ E. coli HST04 dam-/dcm-
E. coli HST04 dam-/dcm-´Â º»·¡ ´ëÀå±ÕÀÌ °¡Áö°í ÀÖ´Â ¸Þƿȭ À¯ÀüÀÚ dam, dcmÀÌ °á¼ÕµÇ¾î ÀÖÀ¸¹Ç·Î, DNA°¡ ´ëÀå±Õ¿¡ ÀÇÇØ ¸ÞƿȭµÇÁö 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. ¶ÇÇÑ, dam, recAÀÇ 2Áß º¯ÀÌ ¼÷ÁÖ´Â »ì¾Æ³²À» ¼ö ¾ø±â ¶§¹®¿¡, º» ¼÷ÁÖ´Â recA+ÁÖ·Î Á¦À۵Ǿú´Ù. ±×·¯¹Ç·Î, ¹Ýº¹¼­¿­À» °®°í ÀÖ´Â ¿Ü·¡ DNA´Â recA À¯ÀüÀÚ¿¡ ÀÇÇØ ÀçÁ¶ÇÕÀÌ ÀϾ °¡´É¼ºÀÌ ÀÖ´Ù. º» Á¦Ç°Àº Ŭ·Î´×¿ë ¼÷Áַδ ÀûÇÕÇÏÁö ¾ÊÀ¸¸ç, ÇüÁúÀüȯ½Ã¿¡´Â, ¹Ì¸® ±¸ÃàµÈ plasmid¸¦ ÃßõÇÑ´Ù.

°íÈ¿À² ¹ü¿ë ¼÷ÁÖ E. coli HB101
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. ¥á- »óº¸¼º ¼±Åà ¼÷ÁÖ E. coli JM109 E. coli JM109´Â pUC°è plasmid vectorÀÇ ÇüÁúÀüȯÀ̳ª M13 phage vector DNAÀÇ ÇüÁúµµÀÔÇÏ´Â °æ¿ì, vector·ÎºÎÅÍ ¹ßÇöÇÏ´Â lacZ ¥á peptide¿Í JM109F¡Ç°¡ ÄÚµåÇÏ´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 ¼­ºêŬ·Î´× À̿ܿ¡ 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
pUC°è plasmid¸¦ ÀÌ¿ëÇÑ À¯ÀüÀÚ library Á¦ÀÛ, ¼­ºêŬ·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¿¡ ÀÇÇØ Å©±â°¡ Å« plasmidÀÇ µµÀÔÀÌ °¡´ÉÇÏ´Ù.

ºÎÀ§ ƯÀÌÀû º¯ÀÌ Ã³¸®¿ë ssDNA Á¦Á¶ ¼÷ÁÖ E. coli CJ236
E. coli CJ236Àº dut, ung º¯ÀÌÁÖÀÇ ´ëÀå±ÕÀ¸·Î dUTPase (Dut)¿Í Uracil-DNA glycosylase (UNG)°¡ °á¼ÕµÇ¾î ÀÖ¾î DNA Áß Thymine (T)ÀÇ ÀϺΰ¡ deoxyuracil (dU)·Î ġȯµÈ DNA¸¦ ÇÕ¼ºÇÑ´Ù. Kunkel¹ýÀº ÀÌ¿Í °°Àº ´ëÀå±ÕÀ» ÀÌ¿ëÇÏ¿© site-directed mutagenesis¸¦ ½Ç½ÃÇÏ´Â ¹æ¹ýÀ¸·Î, º¯À̽Ã۰íÀÚ ÇÏ´Â À¯ÀüÀÚ¸¦ pUC118/119, M13 phage µîÀÇ vector¿¡ Ŭ·Î´×ÇÏ¿© CJ236À» ¼÷ÁÖ·Î ÀÌ¿ëÇÏ¿© ssDNA¸¦ Á¶Á¦ÇϹǷνá Kunkel¹ýÀ¸·Î »ç¿ëÇÒ ¼ö ÀÖ´Â deoxyuracil ÇÔÀ¯ÇÏ´Â DNA¸¦ ¾òÀ» ¼ö ÀÖ´Ù. F¡Ç plasmid (pCJ105)´Â chloramphenicol ³»¼º À¯ÀüÀÚ¸¦ °®°í Àֱ⠶§¹®¿¡ chloramphenicol Á¸ÀçÇÏ¿¡ ¾ÈÁ¤ÇÏ°Ô º¸È£, À¯ÁöµÈ´Ù.

ºÎÀ§ ƯÀÌÀû º¯ÀÌó¸® À¯ÀüÀÚ °íÁ¤ ÁõÆø ¼÷ÁÖ E. coli BMH71-18 mutS
E. coli BMH71-18 mutS´Â mutS ±ÕÁÖ·Î DNA mismatchÀÇ º¹±¸½ÃŰ´Â ´É·ÂÀÌ ÀúÇϵǾî ÀÖ´Ù. µû¶ó¼­ site-directed mutagenesis¸¦ ½Ç½ÃÇÒ ¶§ BMH71-18 mutS¸¦ ¼÷ÁÖ±ÕÀ¸·Î »ç¿ëÇÏ¸é º¯ÀÌ µµÀԺκÐÀÇ ¼öº¹À» ¾ïÁ¦Çϱ⠶§¹®¿¡ µµÀÔÇϰíÀÚ ÇÏ´Â º¯À̸¦ °íÁ¤Çϱ⠽¬¿ö º¯ÀÌ µµÀÔ È¿À²ÀÌ »ó½ÂÇÑ´Ù.

Amber º¯ÀÌ ¼±Åÿë ÇѰ¡´Ú ssDNA Á¦Á¶¿ë ¼÷ÁÖ E. coli MV1184
E. coli MV1184´Â amber suppressor free ±ÕÁÖ·Î amber º¯ÀÌ DNA ¼±º°¿¡ ÀÌ¿ëÇϰųª Mutan-Super Express Km (Code RR022) µî¿¡ ÀÇÇØ º¯À̸¦ µµÀÔÇÒ °æ¿ì, º¯À̰¡ µµÀÔµÈ DNA¸¦ ¼±º°Çϱâ À§ÇØ ÀÌ¿ëÇÑ´Ù. ¶Ç F¡¢ plasmid¸¦ °¡Áö±â ¶§¹®¿¡ M13 phage vector³ª phagemid vectorÀÇ ¼÷ÁÖ·Î ssDNAÀÇ Á¶Á¦¿¡µµ ÀÌ¿ëÇÒ ¼ö ÀÖ´Ù.

Enforcement Cloning System¿ë ¼÷ÁÖ E. coli TH2
E. coli TH2´Â supE-, trpR-, rpsL- ±ÕÁÖ·Î trp promoterÀÇ ÇÏ·ù¿¡ amberº¯À̸¦ °®´Â Ä¡»ç À¯ÀüÀÚ rpsLÀ» ÄÚµåÇÏ´Â Enforcement Cloning System¿ë vector pKF3 DNA¸¦ »ç¿ëÇÏ¿© ¿Ü·¡ À¯ÀüÀÚÀÇ »ðÀÔ À¯¹«¸¦ streptomycin ÷°¡ ¹èÁö¿¡¼­ÀÇ »ýÀ°À¸·Î °£ÆíÇÏ°Ô ÆÇº° °¡´ÉÇÑ ¼÷ÁÖÀÌ´Ù.

¥á- »óº¸¼º ¼±Åà ¼÷ÁÖ E. coli HST02
E. coli HST02´Â ¿Ü·¡ÀÇ ¸ÞƿȭµÈ DNA¸¦ Àý´ÜÇÏ´Â À¯ÀüÀÚ±º ¥Ä(mrrmcrBC-hsdRMS), mcrA¸¦ ¿ÏÀüÈ÷ °á½Ç½ÃÄÑ À¯ÀüÀÚ library Á¦ÀÛ, ¼­ºêŬ·Î´×¿¡ Æø³Ð°Ô »ç¿ëÇÒ ¼ö ÀÖ´Ù. ¶ÇÇÑ F¡¢ plasmid¸¦ °¡Áö°í ÀÖ¾î M13 vector DNAÀÇ ¼÷ÁÖ·Î ÀÌ¿ëÇÒ ¼ö ÀÖ´Ù. pUC°è plasmid ÇüÁúÀüȯ, M13 phage vector DNA ÇüÁúµµÀԽÿ¡´Â ¥â-galactosidaseÀÇ ¥á-»óº¸¼ºÀ» ÀÌ¿ëÇÏ¿© XGal, IPTG¿¡ ÀÇÇØ ÀçÁ¶ÇÕü ¼±º°ÀÌ °¡´ÉÇÏ´Ù.
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 HST04 dam£­/dcm£­

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

E. coli HB101

supE44, ¥Ä(mcrC-mrr), recA13, ara-14, proA2, lacY1, galK2, rpsL20, xyl-5, mtl-1, leuB6, thi-1

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, deoR, recA1, endA1, hsdR17 (rK-, mK+), phoA, supE44, ¥ë-, thi-1, gyrA96, relA1

E. coli CJ236

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

E. coli BMH71-18 mutS

¥Ä(lac-proAB), supE, thi-1, mutS215£º£ºTn10(tetr)/F¡¯[traD36, proAB+, lac I q, lacZ¥ÄM15]

E. coli MV1184

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

E. coli TH2

supE44, hsdS20 (rB-, mB-), recA13, ara-14, proA2, lacY1, galK2, rpsL20, xyl-5, mtl-1, thi-1, trpR624

E. coli HST02

F¡¯ [traD36, proA+B+, lac Iq, lacZ¥ÄM15]/¥Ä(lac-proAB), recA, endA, gyrA96, thi, e14- (mcrA-), supE44, relA, ¥ÄdeoR, ¥Ä(mrr-hsdRMSmcrBC)

ǰÁú

ÇüÁúÀüȯ È¿À²

1 ngÀÇ plasmid DNA¸¦ ÇüÁúÀüȯÇÑ °æ¿ì

100 §¡ E. coli HST08 Premium Competent Cells/1 ng pUC19 plasmid

£¾1¡¿108 transformants/1 §¶ pUC19 plasmid DNA

100 §¡ E. coli HST04 dam£­/dcm£­ Competent Cells/1 ng pUC19 plasmid

£¾1¡¿106 transformants/1 §¶ pUC19 plasmid DNA

100 §¡ E. coli HB101 Competent Cells/1 ng pBR322 plasmid

£¾1¡¿108 transformants/1 §¶ pBR322 plasmid DNA

100 §¡ E. coli JM109 Competent Cells/1 ng pBR322 plasmid

£¾1¡¿108 transformants/1 §¶ pBR322 plasmid DNA

100 §¡ E. coli DH5¥á Competent Cells/1 ng pUC19 plasmid

£¾1¡¿108 transformants/1 §¶ pUC19 plasmid DNA

100 §¡ E. coli CJ236 Competent Cells/1 ng pUC119 plasmid

£¾1¡¿107 transformants/1 §¶ pUC119 plasmid DNA

100 §¡ E. coli BMH71-18 mutS Competent Cells/1 ng pUC119 plasmid

£¾1¡¿107 transformants/1 §¶ pUC119 plasmid DNA

100 §¡ E. coli MV1184 Competent Cells/1 ng pUC119 plasmid

£¾1¡¿107 transformants/1 §¶ pUC119 plasmid DNA

100 §¡ E. coli TH2 Competent Cells/1 ng pBR322 plasmid

£¾1¡¿107 transformants/1 §¶ pBR322 plasmid DNA

100 §¡ E. coli HST02 Competent Cells/1 ng pUC19 plasmid

£¾1¡¿108 transformants/1 §¶ pUC19 plasmid DNA


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