...like rotting eggs but has a longer shelf life when stored properly. Because BME is not as strong as DTT, ..., while expensive, reduce protocol time, can be stored at 4 °C, and are very consistent gel-to-gel.  Ladder...multiple times. It doesn’t degrade during long-term storage and has both high binding capacity and high protein...inexpensive and easily made in the lab. They can be stored at your bench during the procedure and at 4 °C ...Fluorescent proteins are stable, so blots can be stored for weeks to months without loss of signal. While...

...possess resistance genes survive and multiply. Factors contributing to the development of resistance include...www.youtube.com/watch?v=sx1uDYSfINA. When you go to the doctor, it will very likely take time to determine if ...resistant infection or not, because scientists in the laboratory will need to isolate and grow whatever bacteria...method that is frequently used in the clinical laboratory. What can we do to stop the emergence of antibiotic...

...carefully regulated by a mix of positive and negative regulators. The genome of E. coli is about 4.6 million ...bacteria, however, the protein integration host factor (IHF) assists with the process: it bends the DNA...trading out ADP for ATP to re-activate DnaA. These regulatory mechanisms allow E. coli to control chromosomal...nar/gky457 Park, K., Han, E., Paulsson, J., & Chattoraj, D. K. (2001). Origin pairing (‘handcuffing’) ...

... weird black pellet's DNA yielded a new CRISPR effector protein they named Cas7-11. Testing showed that.... Eugene Koonin and Kira Makarova, longtime collaborators of Omar and Jonathan and experts in CRISPR evolution...single protein, shedding the complexity of its ancestors. They classified the system as Class I, Type III-E...Programmable RNA targeting with the single-protein CRISPR effector Cas7-11. Nature. 2021 Sep;597(7878):720-725. doi...

...Ah, the notorious western blot: we meet again. So useful, yet so finicky to design and optimize. Today...due to loading and transfer errors — not to any factors from before the proteins even hit the gel. But ...all, while overloaded proteins may saturate the detector. In between these extremes lies the linear detection...concentration as well as by antibody dilution — both factors we’ve covered in our previous blog posts (1, 2)...

...site of interest can be cloned into an expression vector with Gibson assembly. While this is a comprehensive...is generated by transforming a backbone phagemid vector into an E. coli strain that is deficient in dUTPase... combine pairs of mutations together to test combinatory effects. One-pot saturation mutagenesis The ...2 instances where it does. 1) When the backbone vector has multiple BbvCI sites. If these sites have opposite...

... green fluorescent protein (GFP) from Aequorea victoria and its variants are genetically encoded fluorescent... of fluorescent proteins derived from Aequorea victoria GFP sticks out and can be considered a linker ...Janetopoulos, Chris, Tian Jin, and Peter Devreotes. "Receptor-mediated activation of heterotrimeric G-proteins...et al. "Expanded dynamic range of fluorescent indicators for Ca2+ by circularly permuted yellow fluorescent...

...Split-BioID: A conditional proteomics approach to monitor protein-protein interactions Split-BioID combines... a split BirA* molecule that can reassemble to restore activity when fused to two interacting proteins... in one experiment a scientist can confirm and monitor the interaction of two proteins while also biotin...Split-BioID a conditional proteomics approach to monitor the composition of spatiotemporally defined protein...

...have previously used histone deacetylase (HDAC) inhibitors to increase transcription of the integrated HIV...turned to the CRISPR/Cas9 Synergistic Activation Mediators (SAM) system available from Addgene. This system...TALENs focused primarily on disrupting CCR5, the receptor that HIV-1 uses to enter a cell. ZFNs and CRISPR...W. CRISPR/gRNA-directed synergistic activation mediator (SAM) induces specific, persistent and robust ...

...reduced by adding a protease inhibitor cocktail and phosphatase inhibitors to your lysis buffer. Additionally...ladder runs off the gel. Luckily, this is easily monitored in real time, as the dye front (and many ladders...resources with examples, like BioRad’s Western Blot Doctor page, or robust discussions, like ResearchGate’...longer time Protein degradation Add protease inhibitors to lysate Insufficient denaturing conditions...