1. Transfer 50 ?l of Stbl3 competent cells in a 1.5 ml Eppendorf tube placed on ice . 2. Add between 100ng and 500 ng of plasmid into the tube and resuspend with Stbl3 competent cells (pipet to resuspend, do not use vortex) . 3. Incubate the tube on ice for 30 min, gently resuspend every 10 min. 4.

The results presented here are the first demonstration that E. coli Stbl3 is superior to E. coli Stbl2 in the maintenance of an instability-prone HIV-based plasmid. Based on the presented data, it can be suggested that E. coli Stbl3 might be a host of choice for propagating unstable DNA of lentiviral backbone plasmids. The presented plasmid A wide variety of protocols from Addgene that can be used for basic molecular biology, plasmid cloning, and titering and testing your viral preparations. I transform my ligation into STBL3 competent cells, pick and grow colonies, do minipreps, and check One Shot Stbl3 Chemically Competent E. coli are designed especially for One Shot® Stbl3™ Competent E. coli: 21 vials, 50 µl each Manuals & protocols 28 Jan 2014 This protocol has worked well for us for multiple E. coli strains, including * These strains are not phage resistant. All NEB competent cells are phage resistant. ** Note that this product has a TE of > 1 x 10 8, whereas NEB C2987 has a TE of 1-3 x 10 9 † Note Lucigen 6060-2 has a TE of ? 1x10 8, whereas NEB C2987 has a TE of 1-3x10 9 †† Note Bioline BIO-85025 has a TE of ? 10 7, whereas NEB C2988 has a TE of > 1x10 6 Remember that each of these shortcuts will reduce the efficiency of the transformation, so when higher efficiency is needed follow the complete protocol. Thaw the competent cells in your hand instead of on ice; Reduce step 4 from 20 - 30 mins to 2 mins on ice before heat-shock Modifid protocol for Iarge plasmids (>6 kb) and cDNA libraries : 20-min ice bath followed by 1- min 42?C water bath and another 20-min ice bath. Efficiency will increase 2-5 fold. 4. Further incubation with either