...specific type of virus you are trying to make. For example, bacteriophages will need to be produced in bacteria... P0), infecting new Sf9 cells, and getting an amplified P2 virus. This P2 virus can be produced in a greater...Processing of your baculovirus is straightforward — simply spin down the supernatant in a centrifuge to remove...

...RNA (pegRNA). The pegRNA consists of a target-complementary spacer, a scaffold sequence, and an extension...primer binding site and a reverse transcription template (RTT). The design of the pegRNA has been shown... of different pegRNAs for each desired edit, it simply becomes impossible (or at least unreasonable) to...

...specific epitope that your antibody binds. For example, Myc1-9E10 binds EQKLISEEDL (pronounced “equi-kli-seed-le...be the antibody Mary Poppins!   All of the examples of new versions of Myc1-9E10 mentioned above basically...for the foreseeable future. These newer findings simply add to our understanding of how this antibody works...

... in proximity by the looping of chromatin, the complex of DNA and proteins that forms chromosomes. These... of the CAPTURE method. After protein and DNA complexes are cross-linked, DNA is digested into smaller...and 2) gRNAs Number of loci targeted One Many Sample type(s) Cell lines Cell lines, primary cells ...

... this purpose.Learn about the effects of CO2 on sample pH Cell morphology: Shape and size matter You should...which multiplies every 16 hours). So if you, for example, split cells on a Monday diluting them 1:10, you...contaminated.Learn how Addgene uses barcodes to track samples Keep track of passage numbers Keep an eye on the...

...targeting anywhere in an exon. But things get more complicated if you’d like to use homology directed repair...alternative PAM sequences are longer and often more complex than NGG, resulting in lower overall genome frequency... be less predictable than SpCas9 off-targets. Employing xCas9 for transcriptional regulation and base ...

...want to include a homology directed repair (HDR) template to create knock-ins, point mutations, or to add...transgenic embryos. Cas9-gRNA ribonucleoprotein complexes Purified Cas9 protein and in vitro transcribed...transcribed gRNA are combined to form a Cas9-gRNA complex and delivered to cells using cationic lipids. Transient...

...allow people to, for example, knock out a gene with simultaneous complementation, with a knockout and ...purification; the whole cloning process is so simplistic once you have your parts. That was the motivation...

...getting-started guide for BLAST, which includes a simple explanation of the different BLAST programs, databases...consistently run our blastx searches with the “Low complexity regions” filter unchecked so that these regions...benefit is that blastp searches do not filter low complexity regions by default, meaning that you do not have...

...sequences of guides across 8 CRISPR libraries. Samples are divided into those of African ancestry(orange..., but it still points to a far larger and more complex problem endemic to biomedical research: pervasive...same reference genomes and libraries. Further complicating the issue is the long and ugly history of reporting...

...blots, IP, and affinity purification. Arguably the simplest affinity tag is the polyhistidine (His) tag. Small... a few time-tested tags and fusion proteins as examples. The tables list more common tags than are described...Biotechnology Journal 2012 , (7): 620-634. Full Text A complete list of free handbooks for protein purification...

... array of biological disciplines and scientists employ GFP for a wide number of functions, including: ...importance of the entire GFP protein, which is almost completely required to maintain fluorescent activity; very...modified ES cells, which can then be used for implantation and generation of transgenic mice. Purification...

...to be managing. Being reminded of the seemingly simple actions we can take to help ourselves can be an... in the family to have to miss something – for example, say no to some of those birthday parties. Tell...time. For me, that’s been volunteer work at our temple, my kids’ school, and with professional organizations...

...pBAD-bARGSer-AxeTxe, gas vesicle expression in E. coli is easy: simply grow cells in Lennox LB (10 g/L tryptone, 5 g/L...improved purity and contrast. For in vivo experiments, simply induce injected engineered cells via regular intraperitoneal...a chemical transfection reagent (e.g. PEI-MAX). Simply cotransfect a mixture of 1248 ng of CMV-gvpA-IRES-BFP...

...Eventually I learned that it’s practical because there simply isn’t enough time to read every single paper. I... read the original reference for the data. For example, we summarized how frequently each member of a ...original paper if you wanted to learn more. Another example: we made a schematic of all the proteins in the...

...subunit of DNA gyrase and trapping it in the cleaved complex. This results in DNA breakage and cell death. Conversely...often lost without plasmid addiction systems. For example, Cara Morin and James Kaper used the hok/sok system...K. Burley. "Crystal structure of the MazE/MazF complex: molecular bases of antidote-toxin recognition....

...using viral LTRs as RNAP II promoters is commonly employed in lentiviral and retroviral constructs and we...speaking, promoters in bacteria are less diverse and complex, having fewer parts than those in eukaryotic cells...human ubiquitin C gene Constitutive  As the name implies, this promoter is ubiquitous. human beta actin...

...signaling — think of a mouse with an LED brain implant or a worm wiggling back and forth under blue light...have various disadvantages and limitations – for example, microtubule-destabilizing drugs are impossible... embryo (Aljiboury et al., 2023). It’s a great example of how optogenetics allows you to disrupt and study...

...longer RNAs (such as mRNAs for gene expression). Examples of commonly used RNA pol II promoters for Cas9...cloning of your gRNA of interest. Figure 1: Simplified schematic of agrobacterium-mediated transformation...delivery and expression in planta using chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol...

...convoluted due to researchers sharing and renaming samples throughout history. It was likely isolated in 1918...BL21 strain and derivatives are the most common examples of the E. coli B strain. Common E. coli strains...vectors. Used by Addgene for pooled library amplification. F' mcrA Δ(mcrBC-hsdRMS-mrr) recA1 endA1 gyrA96...