...working with the community to create a fully transparent resource where researchers can easily share, ...

...programmed regulation of DNA replication and gene transcription during vertebrate embryogenesis. My interests...

...(2015): 337-352. PubMed PMID: 26495581. PubMed Central PMCID: PMC4528400. Additional resources on the ...

...  For example, while “PCR test” has recently transitioned from jargon to non-jargon in the context of ...

...molecular cloning consist of: Leader Sequence: Extra base pairs on the 5' end of the primer assist with...

...remove many barriers of participation in terms of travel, costs, and time away from home. You can choose...

...is just 15 minutes a week. Clearly identify constraints and boundaries. Where are the lines of authority...

...Antibodies 101: Flow Cytometry Conventional vs Spectral Flow Cytometry Beyond Surface Labeling in Flow...

...Flow Cytometry Antibodies 101: Conventional vs. Spectral Flow Cytometry Resources on Addgene.org Addgene's...

...Katz: I have a puppy and so I am learning how to train a dog with my daughter. It’s super fun and we are...

...induces the DNA base change without a DNA double-strand break. But base editors aren’t perfect. They may...

...in-house software and also eliminates many of the extraneous ORFs that cluttered our old plasmid maps. Clearly...

...that uses a peer review system. Peer review in traditional journals happens before the manuscript is accepted...

...accuracy across all of our plasmids and provide transparency in sequence discrepancies. While Addgene’s QC...

...staining multiple targets indirectly, you must strategically plan primary/secondary antibody pairing such...

...Pages Other Links Check out Pawluk et al's Video Abstract Read up on All Things CRISPR with These Papers...

...experiment, but a picture of a gel or a sequencing trace file can go a long way in helping us understand ...

...BSA standard concentration versus absorbance. Extrapolate the total protein concentration of the sample...protocol uses a dry transfer device but can be adapted for wet and semi-dry transfer approaches. Last Update...blocking buffer and optimal antibody concentrations. Consider titrating your antibody to determine the optimal...protein concentration and prepare the lysate for SDS-PAGE Determine the protein concentration using a ...well. Assemble the transfer sandwich as follows: Unseal the iBlot 2 PVDF Mini transfer stack. Set the Top...iBlot 2 Gel Transfer Device. Wet the pre-run gel in deionized water and place it on the transfer membrane... parameters are correct. Transfer for 5–6 min for proteins <30 kDa. Transfer for 8–10 min for proteins...

...-term protein expression observed using transient transfection approaches, generating cell lines using...experiments in a stable cell line, as opposed to transiently-transfected cells, increases reproducibility, as it... the variation associated with repeated transient transfection. Some lentiviral vectors deliver mammalian...stable cell culture after transduction. Performing antibiotic selection on transduced cells enables elimination... the final concentration of polybrene in each well should be 10 µg/mL. Pro-Tip Transducing too many cells... may have 1 copy of the transgene, while others have >1 copy of the transgene in their genome. This can...high levels of the transgene may have reduced growth rates, especially if that transgene is toxic. Eventually...

...Polymerase III transcription for generation of shRNA transcripts. cPPT Central polypurine tract, cPPT, improves...a Qiaquick gel extraction kit. Elute in 30 μL of ddH 2 O. Measure the DNA concentration. C.4 Ligating ...HIV-derivative viruses. For transient knockdown of protein expression, you may transfect plasmid DNA directly...Day 2: b. Perform the transfection in the late afternoon because the transfection mix should only be incubated...Digesting pLKO.1 TRC-Cloning Vector C.4 Ligating and transforming into bacteria D. Screening for Inserts D.1 Recommended...materials F.2 Determining the optimal puromycin concentration F.3 Protocol for lentiviral infection and selection...pLKO.1 can be introduced into cells via direct transfection, or can be converted into lentiviral particles...