...MaxiPreps (Qiagen HiSpeed Max, Catalog #12663) Tips (1000 µL, 200 µL, 10 µL) Bacti Cell Spreaders (VWR, 60828... evenly with a sterile spreader until all liquid has been absorbed by the agar. This usually takes 1-2... 100 x 100 ÷ 0.1 This number should be at least 1000X greater than number of perturbations in library....least 10 colonies on the most diluted plate for 1000x coverage of a library of 100,000 plasmids. Colonies...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette...sealer and press the ‘Seal’ button. After the plate has been sealed, proceed to thermocycling. Thermal Cycling...

.... Cas9 nickase (Cas9n), a D10A mutant of SpCas9, has one active nuclease domain and one inactivated one... available. SaCas9 (from Staphylococcus aureus ) has a coding sequence ~1 kb shorter than SpCas9 while... SpCas9. FrCas9 (from Faecalibaculum rodentium ) has increased fidelity, and recognizes NNTA, a PAM sequence... create the adenine base editor. The TadA domain has also been further evolved to catalyze cytidine deamination...editing requires more optimization and typically has lower efficiency (< 50%) than base editing (> 90%...modifications to eliminate target gene function. It has also been used extensively to screen for novel genes... those for yeast or drosophila . Although CRISPR has been less widely used in bacteria due to technical...

...an impure AAV preparation. The 15% iodixanol step has 1M NaCl to destabilize ionic interactions between... and forth a few times to mix the iodixanol that has settled at the bottom of the column. We recommend...bring up the volume with formulation buffer. Use a P1000 to the bottom of the filter and pipette up/down ...

...antibiotic required for effective over-agar selection has been empirically determined. See selection curve ...without antibiotic High concentration (100 mg/mL, 1000x) carbenicillin stock solution in sterile water (...