...Incubate plates at 37°C overnight. Tips and FAQ How can I save time when carrying out transformations? If you...is a good idea to outgrow for at least 20-30 mins) I get very few if any transformants when transforming...transforming large plasmids (>10 kb) or BACs, what can I do? Chemically competent cells are fast and easy to use... Follow the manufacturer's instructions for each. I got no transformants. What went wrong? Check that ...learn how to isolate single bacterial colonies. Equipment Shaking incubator at 37 °C Stationary incubator...

...Solution I - Resuspension Buffer 25 mM Tris-HCl (pH 8) 50 mM glucose 10 mM EDTA Store Solution I at 4°C .... Resuspend the pellet in 100 μL of cold Solution I. Vortex the solution for 2 min or until all bacteria...bottom of this page. Last Update: Feb. 8, 2018 Equipment Desktop microcentrifuge Desktop vortexer Vacuum...

...Pooled Libraries Molecular Biology Reference How do I process my Addgene pooled library? Introduction Please...total plasmid DNA. Tips and Troubleshooting What do I do if my transformation efficiency is not high enough...light in the cuvette during the electroporation step. I got a heavy recombined band by digest. Recombination...ensure that outgrowth and competition are limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser...and follow all specifications described in the equipment manual. Immediately add 1 mL SOC to cuvette. Remove...

...linearized. Treat the purified AAV samples with DNase I to eliminate any contaminating plasmid DNA carried...Haase M, Muether N, Hausl M, Rauschhuber C, Huber I, Nitschko H, Busch U, Sing A, Ehrhardt A, Baiker A...set-up: 2 h qPCR run: 1.5 h Data analysis: 30 min Equipment qPCR instrument Heating plate Pipettors 1–10 µL...

...ligation. Use T4 DNA Polymerase or Klenow DNA Polymerase I for 3′ overhang removal and 5′ overhang fill-in. If...how to analyze your restriction digest results. Equipment Electrophoresis chamber Pipetman Reagents Liquid...

...your gene of interest Triton X-100 Benzonase/DNAse I (Millipore 71205-3) 40% Polyethylene Glycol 8000 (...am): Transfect cells Day 7 (am): Harvest cells Equipment Class II, Type A2 Biological Safety Cabinet 0.5...