...titrating each lot of AAV in vivo to determine the optimal dosage. At Addgene, we don’t reserve lots, but ...

...also the proposed salary range) should help. In smaller companies, technical support is often covered by...

...transcriptase (RT) variants, PE6a and PE6b are 500–800 bp smaller than previous editors, but just as effective and...

...demonstrating it. Requests like this are both very normal and very helpful; don’t be afraid to ask! Especially...

...-Kulka, and Gad Glaser. "An Escherichia coli chromosomal" addiction module" regulated by guanosine [corrected...

...complex challenges, quickly breaking them up into smaller achievable tasks, and then executing on them in...

...Substantive conflict occurs when people or teams who normally get along well with each other disagree about ...

...fragments of linearized DNA migrate slower than smaller linearized fragments. You can separate your backbone...

...nominations for government fellowships. The closing and formal evaluation of the pilot program IMP 2014/2015, ...

...yourself and the world around you, it’s perfectly normal to realize that you have new interests and that...

...Cell-based (in vitro) Can be used on tissue from whole animal models, no exogenous component required (e.g. dsODN...

...show efficient absorption at 650 nm, APC-Cy7's optimal excitation happens at higher wavelengths of about...

...management consulting firms), sponsor more H-1Bs than small startups and firms with a non-international focus...

... of cold 4% paraformaldehyde in PBS on ice for 15 min . Pro-Tip While 4% Paraformaldehyde fixation works...labeling HeLa cells for a target protein using the formaldehyde fixation method. The protocol may need to be...coated coverslips HeLa cells 24-well plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI...be better for some applications. Remove the paraformaldehyde and follow your institution's laboratory safety...concentration in antibody dilution buffer. Pro-Tip The optimal antibody concentration will vary but generally ...fluorescent filters. Tips and Troubleshooting The optimal fixation method will vary depending on the sample...

...Web resources Addgene’s mammalian RNAi website: www.addgene.org/mammalianrnai The RNAi Consortium (TRC...Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal 21-mer targets in your gene B.2 Order oligos compatible... F.1 Recommended materials F.2 Determining the optimal puromycin concentration F.3 Protocol for lentiviral...resistance gene for selection of pLKO.1 plasmid in mammalian cells. sin 3’LTR 3’ Self-inactivating long terminal...’s lab. For more information, visit Addgene’s Mammalian RNAi Tools page. Several other laboratories have...Designing shRNA Oligos for pLKO.1 B.1 Determining the Optimal 21-mer Targets in your Gene Selection of suitable...Weinberg lab. For more information, visit Addgene’s Mammalian RNAi Tools page. E.2 Protocol for Producing Lentiviral...

...represents the first decimal place. For the P20, this represents the second decimal place, and for the ...pipette in many scenarios to accurately dispense small amounts of liquid (think: 0.1 µL to 1 mL). When ...volume display on these pipettes will also have small tick marks at the bottom. For the P1000, this represents...the P2 this represents the third decimal place. Procedure How to Pipette Now that you know the different...pipetting from a relatively large container holding a small volume of liquid, tilt the container holding the...accurately and precisely is important since even small changes in the volume could affect your experiments...

...buffer and optimal antibody concentrations. Consider titrating your antibody to determine the optimal dose.... concentration in blocking buffer. Pro-Tip The optimal concentration will vary between antibodies but ...detect the bands. Tips and Troubleshooting The optimal lysis buffer will vary depending on the sample ... buffers to find the best for your target. The optimal concentration and blocking buffer will vary between...consider titrating your antibody to determine the optimal dose. To ensure that your antibody is both functioning...

...overlapping DNA fragments can be joined in a single isothermal reaction. With the activities of three different...two-part Gibson reaction if you're only making a small change in a plasmid (such as point mutations). Generate...concentrations. The Gibson Assembly master mix (isothermal reaction mix) consists of three different enzymes...-Stranded DNA-Binding protein (ET SSB) to the isothermal reaction mix. ET SSB protects 3’ ssDNA ends from... in the Gibson reaction are different than with normal PCR primers (Gibson et al. , 2010). Number of fragments...Cepko C. (2020). A Simple Enhancement for Gibson Isothermal Assembly. bioRxiv . https://doi.org/10.1101/2020.06.14.150979...

...your substance maximally absorbs. In the case of nucleic acid (DNA and RNA), the maximal absorbance is ...and most have a specific wavelength that they maximally absorb. By measuring the absorbance of a liquid...is at 260nm. Protein maximally absorbs at 280nm and the ratio of nucleic acid to protein (260/280) is generally...days, many labs have a NanoDrop, which is a very small spectrophotometer that can accurately read DNA concentration...

...While simple homogenization is effective for most mammalian tissues; more hardy tissues such as bone, or bacteria...Pro-Tips If you anticipate your RNA yield to be small, RNase-free Glycogen may be used as a carrier to...cycles of your entire RNA sample, consider making smaller aliquots of it and storing those in -80°C. Option...cycles of your entire RNA sample, consider making smaller aliquots of it and storing those in -80°C. Solution...