...frequency and exact sequence of targeted alleles. Involves PCR amplification of targeted region from DNA,...

...CRISPRs into Cells of Interest NOTE: This protocol involves the delivery of CRISPR/Cas9 plasmids by electroporation...

...of interest. pJT106b3 31393 Red light inducible inverter Red light (650 nm) Christopher Voigt When combined...

...Karpova and Schaffer labs. See the Retrograde AAV Inventory Page . Don’t See What You’re Looking For? Our ...

...window) Qiagen , (Link opens in a new window) Invitrogen , and (Link opens in a new window) Promega sell...fully resuspended. Add 200 μL of Solution II and invert the tube carefully 5 times to mix the contents.... μL of cold Solution III to each tube. Mix by inverting several times. A white precipitate will be formed...reactions. Air dry the pellet (can be done by inverting the tube at an angle over kimwipe) for 20-30 min...ethanol and 1/10 volume of 3 M Na-acetate (pH 4.8). Invert the microfuge tube to mix. Optional: Place the ... Pour out the supernatant in the sink. Open and invert the tubes on a paper towel to drain them out. Wash...about losing the pellet. Dry with vacuum or by inverting over paper towel for 5-20 min. Resuspend dry DNA...

...Addgene's viral vectors See Addgene's Current AAV inventory , which includes all viral vectors transferred...

... 1mm thick, 10-well, Invitrogen NP0321BOX 20X MOPS SDS running buffer, Invitrogen NP0001 4X NuPage sample...buffer, Invitrogen NP0007 10X NuPage sample reducing agent (500 mM dithiothreitol, DTT), Invitrogen NP0009...NP0009 PageRuler Plus Prestained protein ladder, Invitrogen 26619 Pipette tips, 10 µL, VWR 76322-132 Pipette...MOPS buffer into 475 mL of deionized water. Gently invert to mix. Procedure Section 1: SDS-PAGE Add 5 µL ...

...Tween-20 to into 999.5 mL 1X PBS Cap the bottle and invert several times to mix. Carefully remove the plate...250 mg of BSA to 25 mL of PBS. Cap the tube and invert several times to mix. Using a multichannel pipette...Dilute 50 mg BSA into 50 mL PBS . Cap the tube and invert several times to mix. Dilute the primary antibody...three concentrations between 1–10 µg/mL. Cap and invert the tubes several times to mix. Using a multichannel...concentrations in the antibody dilution buffer. Cap and invert the tube several times to mix. Using a multichannel...Solution from the TMB substrate kit. Cap the tube and invert several times to mix. Using a multichannel pipette...

...serum-free media Invitrogen: #31985 Dulbecco’s Modified Eagle Medium (DMEM) Invitrogen: #11995 Fetal Bovine...Bovine Serum (FBS) Invitrogen: #16000 Penicillin/Streptomycin Invitrogen: #15140-122 Polypropylene tubes... buffer NEB #B0202S DH5 alpha competent cells Invitrogen #18258-012 Qiaquick gel extraction kit Qiagen...homologous recombination of unstable regions, such as Invitrogen Stbl3™ or NEB Stable cells. This will ensure ...

...fluorophores used is important. It is best to inversely pair the brightness of the fluorophore with the...

...incubator Pipet controller Hazardous waste container Inverted fluorescence microscope Reagents DMEM high glucose...mL of DMEM complete. Mix well by pipetting or inverting. Aliquot 2 mL of cell suspension into each well...

...your application. A diagnostic digest typically involves ∼500 ng of DNA, while molecular cloning often ...s) following the digestion reaction. This may involve incubating the reaction at 70 °C for 15 mins, or...

... thoroughly by inversion. Add 1 mL water-saturated phenol, mix thoroughly by inversion. Add 0.2 mL of ...

...spreading of antibiotic makes it easy for an investigator to conveniently plate and select transformed...

...fusion proteins, point mutations, deletions, etc.) involves selecting one or more unique oligonucleotide primers...

...depending on the model of water bath, but will likely involve a dial or a digital display you can set to the ...

...centrifuge. Immediately stop the centrifuge and investigate the issue. When you are finished spinning your...

...propagation. The majority of ligation reactions involve DNA fragments that have been generated by restriction...

...gene. This trick can also enable replacement of "inverse PCR" reactions with a two-part Gibson reaction ...

... 10 μg/mL polybrene. Mix well by pipetting or inverting. Aliquot 1.35 mL of the cell suspension into each...