产品介绍

基 因 型F-ompT hsdSB(rB-mB-)gal dcm(DE3) pRARE(argU，argW，ileX，glyT，leuW，proL)(CamR)简 要 说 明High5TM类型Rosetta(DE3) 体验态内部拥有氯霉素抗性，提供结肠杆菌缺乏性的6 种希有账户密码子(AUA，AGG， AGA， CUA，CCC， GGA) 使用的tRNA，提高自己外源表观遗传，还是比较是真核表观遗传在原核系统的中的把你想理解出来的水平，该菌株染色的体梳理了λ噬菌体DE3区（DE3区所含T7噬菌体RNA配位聚合反应物酶），一并把你想理解出来T7 RNA配位聚合反应物酶和结肠杆菌RNA配位聚合反应物酶，该用做pET类型，pGEX，pMAL等质粒的淀粉酶把你想理解出来。High5TM类型Rosetta(DE3) 体验态内部由特俗施工工艺定制，经pUC19质粒验测转变利用率可高达108cfu/μg。操 作 说 明1.取100μl冰面运动消融的Rosetta(DE3)体验态内部，放入原因质粒并缓缓混匀，冰面运动静置2两几15秒钟。2.42℃水浴热激45秒，飞速放回冰面运动并静置2几15秒钟，摆动会降低转变利用率。3.向离心法法管上放入700μl含有消炎药类的灭菌培植基（2YT或LB），混匀后37℃，200rpm溶栓60几15秒钟。4.5000rpm离心法法一几15秒钟收菌，留取100μl左右两上清缓缓吹打重悬菌块并涂抹到含相关联消炎药类的2YT 或LB培植基上。5.将pad反过来放于37℃培植箱包夜培植。注 意 事 项1.体验态内部好一点在冰面运动消融。2.掺入质粒时应当温柔实操。3.转变高浓硫酸氧化还原电位的质粒可相关联减轻最后用做涂板的菌量。4.引发时，IPTG浓硫酸氧化还原电位可选择（0.1-2mM均可）。5.为拿到须要量的淀粉酶，最好引发时，湿度，IPTG浓硫酸氧化还原电位需实验性者提高。Sample Induction Protocol (for reference only)1.Inoculate a single colony from a freshly streaked plate into 5 ml of LB medium containing the appropriate antibiotic for the plasmid and host strain.2.Incubate with shaking at 200 rpm at 37℃ overnight. 3.Inoculate 50 ml of LB medium containing the appropriate antibiotic with 0.5 ml of the overnight culture prepared in step 2(use the 500 ml triangular flask as the container would be better).4.Incubate with shaking at 150 rpm at 37℃ until the OD 600 reaches 0.5-0.8.5.(Optional)Pipet 1ml of  the cultures into clean microcentrifuge tubes and  place the tubes on ice until needed  for gel analysis or storage at  -20℃. These will serve as the non-induced control samples.     6.Add IPTG to a final concentration of 1 mM.  Optimal time for induction of the target protein may vary from 2-16 hours, depending on the protein.7.Incubate with shaking at 120 rpm at 37℃ for 3-4 hours. To determine the optimal time for induction of the target protein, it is recommended that a time course experiment be performed varying the induction from 2-16 hours.8.Place the culture on ice for 10 minutes. Harvest cells by centrifugation at 5,000 x g for 101.minutes at 4℃.9.Remove the supernatant and store the cell pellet at -20℃ (storage at lower temperatures is also acceptable). IPTGPrepare a 1 M solution of IPTG (Isopropyl-β-D-thiogalactoside; Isopropyl-β-D-thiogalactopyranoside) bydissolving 2.38 g of IPTG in dd water and adjust the final volume to 10 ml. Filter sterilize before use.

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