TB1 Chemically Competent Cell说明书产品货号: ML-G2025
保存条件: -80℃
产品规格: 10×100μl 50×100μl
产品介绍基 因 型F– ara Δ(lac-proAB) rpsL (Strr)[φ80 dlacΔ(lacZ)M15] thi hsdR简 要 说 明TB1感受态细胞来源于JM 83,是JM 83 hsdR型,只含有大肠杆菌RNA polymerase,缺少 BL21 (DE3)菌株的 T7 RNA polymerase,适合NEB公司的pMAL 系列质粒原核蛋白表达(The pMAL vectors use the E. coli RNA polymerase),不能用于pET系列质粒的表达,具有链霉素抗性。MLBio High5TM系列TB1感受态细胞由特殊工艺制作,经pUC19质粒检测转化效率高达108cfu/μg。操 作 说 明
1.取100μl冰上融化的TB1感受态细胞,加入目的质粒并轻轻混匀,冰上静置25分钟。2.42℃水浴热激45秒,迅速放回冰上并静置2分钟,晃动会降低转化效率。3.向离心管中加入700μl不含抗生素的无菌培养基(2YT或LB),混匀后37℃,200rpm复苏60分钟。4.5000rpm离心一分钟收菌,留取100μl左右上清轻轻吹打重悬菌块并涂布到含相应抗生素的2YT或LB培养基上。5.将平板倒置放于37℃培养箱过夜培养。注 意 事 项1.MLBio 感受态细胞最好在冰上融化。2.混入质粒时应轻柔操作。3.转化高浓度的质粒可相应减少最终用于涂板的菌量。4.诱导时,IPTG浓度可选(0.1-2mM均可)。5.为获得需要量的蛋白,最佳诱导时间,温度,IPTG浓度需实验者优化。
Sample Induction Protocol (for reference only)1. Inoculate a single colony from a freshly streaked plate into 3ml 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. (0.6 recommended; about 2.5h).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 2-4 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 10minutes 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.
产品标签
用户评论(共6条评论)
- Rosetta-gami 2(DE3)
- ArcticExpress (DE3) pRARE2感受态细胞
- Rosetta-gami(DE3)pLysS感受态细胞
- BL21 电击感受态细胞
- Tuner(DE3)感受态细胞
- OrigamiB(DE3)感受态细胞
- BL21-CodonPlus (DE3)-RIPL感受态细胞
- BL21 Star(DE3)pLysS感受态细胞
- Rosetta-gami B(DE3)
- M15(pREP4) 感受态细胞
- Rosetta(DE3) 感受态细胞
- BL21(DE3)pLysS感受态细胞
- BL21(DE3) 感受态细胞
- Tuner(DE3)pLysS感受态细胞
- OrigamiB(DE3)pLysS感受态细胞
- ER2566 感受态细胞
- BL21感受态细胞
- Rosetta 2(DE3)感受态细胞
- C43(DE3) pLysS感受态细胞
- BL21 Star(DE3)感受态细胞
- BL21(AI)感受态细胞
- OverExpress C43(DE3)电击感受态细胞
- OverExpress C43(DE3)感受态细胞
上海酶联生物科技有限公司
Shanghai Enzyme-linked Biotechnology Co., Ltd.
订购热线:4008-898-798 / 021-54720761
传 真:021-54222852
地址:上海市松江区云凯路66号T2楼1603室
酶联官方手机二维码
微信扫描二维码,参与酶联活动,
劲爆优惠触手可得!
本网站销售的所有产品仅用于工业应用或者科学研究等非医疗目的,不可用于人类或动物的临床诊断或治疗,非药用,非食用。