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View additional product information for MultiShot™ StripWell Mach1™ T1 Phage-Resistant Chemically Competent E. coli - FAQs (C869601)
9 product FAQs found
我们推荐使用mcr/mrr–菌株，这种菌株可以防止甲基化的真核DNA在大肠杆菌宿主中的扩增受到限制。我们还推荐使用T1R菌株，因为T1是基因组/cDNA文库中一种常见的污染。
我们的Mach1-T1R感受态细胞比我们其它常规克隆菌株长得快。它的倍增时间是54分钟，而普通的克隆菌株如DH5α的倍增时间则需要增加70分钟。37摄氏度下将转化混合液铺板后8小时左右就可以在培养板上看到Mach1-T1R菌株的克隆。将过夜培养的单个大克隆接种到1.5毫升离心管37摄氏度下培养4小时就能进行质粒小抽。
We would recommend a mcr/mrr- strain, which prevents restriction of methylated eukaryotic DNA in the E. coli host. We would also recommend using a T1R strain, as T1 is a common contaminant in genomic/cDNA libraries.
Our Mach1-T1R competent cells grow faster than any of our common cloning strains. It has a doubling time of 54 minutes versus doubling times in excess of 70 mins for standard cloning strains, such as DH5α cells. Colonies of Mach1-T1R begin to be visible on a plate 8 hours after plating the transformation mix at 37 degrees C. It can be mini-prepped from 1.5 mL cultures in as little as 4 hours at 37 degrees C after inoculation with a single large overnight colony.
There are situations where either because of time or process limitations, elimination of the heat shock and recovery time post-heat shock in the transformation protocol would be desirable. Testing was performed on many of our chemically competent cells in both the MultiShot StripWell and MultiShot FlexPlate format with a shortened transformation protocol. Use of a rapid transformation protocol (add DNA, no heat shock, no recovery period, plate on warm agar) yielded up to a log lower in transformation efficiency using the control DNA. This rapid transformation protocol can be performed with expectation of lower colony counts than our standard high efficiency transformation protocol and can only be performed with plasmids that use ampicillin as their antibiotic resistance. The rapid protocol is ideal when using a liquid handling system for high-throughput transformations with high cloning efficiency. Please see the product manual for full details on this rapid transformation protocol option.
The MultiShot FlexPlate is a PCR plate and can be placed in a thermal cycler or heat block set at 42 degrees C to transform the cells by the heat shock method. The duration of the heat shock performed in these devices has yielded similar transformation efficiencies as that seen with heat shock done in a water bath.
We offer the following E. coli strains in the MultiShot StripWell format: TOP10, DH5alpha T1R, Mach1 T1R, Stbl3, and BL21 Star (DE3) chemically competent cells.
We offer the following E. coli strains in the MultiShot FlexPlate format: TOP10, DH5alpha T1R, DH10B T1R, Mach1 T1R, OmniMAX2 T1R, and Stbl3 chemically competent cells.
The MultiShot StripWell format is supplied as pre-aliquoted competent cells in 96 tubes contained in a rack. The tubes are attached in strips of 8 tubes, and there are 12 strips that contain 50 µL of chemically competent cells. This format offers the most flexibility to transform 1 tube or up to all 96 tubes at once and is intended for varied or medium-throughput use.
The MultiShot FlexPlate format is supplied as pre-aliquoted competent cells in a 96-well PCR plate. Each well contains 20 µL of competent cells. The plate can be separated at each column of 8 wells, offering a high level of flexibility to utilize various increments of 8 wells per transformation. The wells of the plate are heat sealed, preventing detachment of the aluminum seal in the -80 degrees C freezer and preventing contamination. This format is compatible with automated liquid handling platforms and is intended for medium- to high-throughput use.
The MultiShot format is supplied as pre-aliquoted competent cells in 5, 96-well thin-wall polycarbonate microplates. Each well of the 96-well microplate contains 15 µL of chemically competent cells. This format is compatible with automated liquid handling platforms and is intended for high-throughput use.