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Stellar¢â Competent Cells
³ôÀº ÇüÁúÀüȯ È¿À²À» °¡Áø E. coli competent cell·Î, ¿Ü·¡ÀÇ ¸Þƿȭ µÈ DNA¸¦ ºÐÇØÇÏ´Â À¯ÀüÀÚ (mrr, hsdRMS, mcrBC, mcrA)°¡ °á¼ÕµÇ¾î ÀÖ´Ù. ÀÌ ¼¼Æ÷´Â ±æÀ̰¡ ±ä cDNA library¿Í genomic library Á¦ÀÛ, subcloning, ¸Þƿȭ µÈ DNAÀÇ cloning µî ´Ù¾çÇÑ ½ÇÇè¿¡ »ç¿ëµÉ ¼ö ÀÖ´Ù. ¶ÇÇÑ, LacZ alpha À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÑ vector·Î ÇüÁúÀüȯ ½Ã blue/white screening¿¡ »ç¿ëÇÒ ¼ö ÀÖ´Ù. ÀÌ Á¦Ç°Àº In-Fusion¢ç Snap Assembly Master Mix (Code 638947)¿Í ÇÔ²² »ç¿ëÇϸé ÃÖ°íÀÇ cloning È¿À²À» º¸ÀδÙ.

Stellar¢â Competent Cells (dam-/dcm-)
DNAÀÇ ¸Þƿȭ¿¡ ÇÊ¿äÇÑ À¯ÀüÀÚ (dam, dcm)°¡ °á¼Õ µÈ E. coli competent cellÀÌ´Ù. ¿©±â¼­ »ý»êµÈ plasmid´Â dam ¶Ç´Â dcm ¸Þƿȭ·Î Àû¿ëÀÌ ¾î·Á¿ü´ø Á¦ÇÑÈ¿¼Ò¸¦ »ç¿ëÇÒ ¼ö ÀÖ´Ù. dam À¯ÀüÀÚ¿Í recA°¡ °°ÀÌ °á¼Õ µÇ´Â °æ¿ì ¼¼Æ÷°¡ »ç¸êµÇ±â ¶§¹®¿¡ ÀÌ ¼¼Æ÷´Â recA À¯ÀüÀÚ¸¦ ¹ßÇöÇϰí ÀÖ´Ù. À̶§¹®¿¡ cloning¿¡¼­ host cell·Î »ç¿ëÇÒ °æ¿ì ¹Ýº¹ ¼­¿­À» °¡Áö´Â ¿Ü·¡ÀÇ DNA°¡ recA À¯ÀüÀÚ¿¡ ÀÇÇØ º¯À̰¡ ÀϾ ¼ö ÀÖ´Ù. µû¶ó¼­ À§¿¡¼­ ¾ð±ÞµÈ plasmid Á¦ÀÛ¿¡¸¸ »ç¿ëÇÏ´Â °ÍÀ» ÃßõÇÑ´Ù.

Stellar¢â Electrocompetent Cells
ElectroporationÀ» »ç¿ëÇÑ ÇüÁúÀüȯÀ» À§ÇÑ Stellar¢â Competent CellsÀÌ´Ù.
Competent cells Ư¡

Strain

Ư¡

Stellar¢â

Transformation efficiency

> 5 ¡¿ 108 cfu/§¶ pUC19

Genotype

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

Stellar¢â (dam-/dcm-)

Transformation efficiency

> 1 ¡¿ 106 cfu/§¶ pUC19

Genotype

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

Stellar¢â Electro

Transformation efficiency

> 1 ¡¿ 109 cfu/§¶ pUC19

Genotype

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

¿ëµµ
- Cloning ¹× subcloning
- ¸ÞƿȭµÈ DNA cloning
- cDNA¿Í genomic library Á¦ÀÛ
- ±æÀ̰¡ ±ä cDNA library Á¦ÀÛ
- dam ¶Ç´Â dcm ¸Þƿȭ°¡ µÇÁö ¾ÊÀº plasmid Á¦ÀÛ (dam-/dcm-)
º¸Á¸
- Competent cell: -70¡É
- ±× ¿Ü: -20¡É