One Shot™ Mach1™ T1 噬菌体耐受化学感受态大肠杆菌

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 other strains available that may function similarly to Stbl2 cells in stabilizing inserts or vectors with repeated DNA sequences. However, one advantage of Stbl2 cells over many similar strains is that they are sensitive to Kanamycin, so you can use Stbl2 to propagate plasmids containing a Kanamycin resistance marker.