产品介绍

基 因 型E. coli B F- dcm ompT hsdS(rB- mB-) gal [malB+]K-12(λS)简 要 说 明BL21是早期研发的使采用原核抒发方式的菌株，BL21（DE3）、Rosetta、OrigamiB(DE3) 等这些的原核抒发方式菌株均渠道于BL21菌株。该菌株常见使采用非渗透性蛋清的抒发方式，可含T7 RNA缩聚酶，全部不可能使采用由T7重新启动子带动的蛋清抒发方式(如：pET编的的)；但具有刺激性结肠杆菌RNA缩聚酶，能够使采用tac或trc等运用结肠杆菌RNA缩聚酶的原核系统的的抒发方式（如：pGEX，pMAL质粒）。High5TM编的的 BL21感言态生殖细胞膜核由特有技艺生产制作，经pUC19质粒论文检测转成利用率达107cfu/μg。操 作 说 明1.取100μl雪上的溶化的BL21感言态生殖细胞膜核，融入意义质粒并柔美地混匀，雪上静置2分之五钟。2.42℃水浴热激90秒，快速放回雪上并静置2一20分以内，摇动会调低转成利用率。3.向离心力法管上融入700μl可含消炎药的无菌室养育基（2YT或LB），混匀后37℃，200rpm溶栓60一20分以内。4.5000rpm离心力法一一20分以内收菌，留取100μl以内上清柔美地吹打重悬菌块并涂覆到含相对应的消炎药的2YT或LB养育基上。5.将平面上下颠倒放于37℃养育箱過夜养育。注 意 事 项1.感言态生殖细胞膜核最优在雪上的溶化。2.混有质粒时该柔美方法。3.转成高溶液氧化还原电位的质粒可相对应的减掉后面使采用涂板的菌量。4.诱骗型时，IPTG溶液氧化还原电位可供选择（0.1-2mM均可）。5.为获得了需量的蛋清，最优诱骗型时间间隔，温暖，IPTG溶液氧化还原电位需实验设计者简化。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. 

此产品需要干冰运输发货，会根据路途及包装大小收取一定的干冰运费。