...Silver.  Most types of biological motion (whether endocytosis, vesicle trafficking, or muscle contractions)...video. Most types of biological motion (whether endocytosis, vesicle trafficking, or muscle contractions)...studied for many decades, and they were known to be resistant to detergents and degradation under a variety ...demonstrated that R bodies can be “tuned” with mutagenesis to extend at different pH values. Using a screen...in a particular region of the R-body. Further mutatgenesis of this region produced R-bodies that could ...Organization and expression of genetic determinants for synthesis and assembly of type 51 R bodies. J. Bacteriol...

...for experience in “statistical analysis”, mention “statistical analysis” (word by word) in your resume...large data sets and are familiar with statistical analysis. If you have done fluorescent imaging, then you...you probably have experience using image analysis software. If you use computer programs to predict protein...universities even have officers working specifically to assist graduate students in job searching. Start your ...resume writing, as well as additional videos with emphasis for graduate student resumes UC Irvine Non Academic...

...mitochondrial homeostasis maintained through mechanisms like fission, fusion, biogenesis, and mitophagy...proteins would be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that...photostability is crucial. These properties ensure a more consistent and stable signal over extended periods, which...clear differentiation between multiple dyes during analysis. Image resolution is impacted by several factors...fluorescence. When it comes to protein labeling and analysis, dyes should be small enough not to hinder protein...focuses on commercializing their cell and protein analysis products. You can find plasmids deposited by Promega...

...biology in generalized pustular psoriasis Generalized pustular psoriasis is a chronic autoinflammatory ...says Iyer. Since the RESOLUTE plasmid collection consists of human and murine solute channels, the team ...in the biology of GPCRs and G proteins, with an emphasis on oligomerization, signalling complex assembly...nucleus,” says Dr. Terry Hébert, a professor and assistant dean of biomedical science education at McGill... University.  These protein-receptor complexes consist of many parts that work together on the cell membrane...

...workflow and its analysis can be found here: A Novel Next-Generation Sequencing and Analysis Platform to Assess... example, the CAG promoter, a hybrid promoter consisting of a CMV enhancer, chicken beta actin promoter...the FASTQ data from our instrument and begin the analysis. We use a modified version of the Python program...Ersing I, Haery L, Harten DeMaio K, Sanders E, Tasissa M, Kostman M, Tillgren M, Makana Hanley L, Mueller... (2020) A Novel Next-Generation Sequencing and Analysis Platform to Assess the Identity of Recombinant...

...the infants. Almost all patients have achieved persistent immunological reconstitution with a normally ...disease-associated mutations. AAV vectors maintain persistent transgene expression over many years in postmitotic...limited to hemophilia B, LPL deficiency, Cystic fibrosis, Muscular Dystrophy, Parkinson’s disease, and ... form of blindness known as Leber’s congenital amurosis (LCA). This disease is caused by mutations in ... the human gene RPE65, patients with clinical diagnosis of LCA and mutations in RPE65 (n=3), showed significant...Furthermore, deeper understanding of disease pathogenesis will enable robust therapeutic outcomes. In ...

...carbohydrates that can inhibit the cell lysis process. To ensure that lysis is complete, use a reagent like Qiagen...your plasmid yield is sub-optimal? If you are consistently getting sub-optimal plasmid yields from your...Addgene uses and recommends DH5𝝰! This strain consistently gives good, high quality preps due to its endA1...Qiagen’s LyseBlue, which uses color to show you if lysis is incomplete. Protein expression strains are not...produce fewer transcripts of their antibiotic resistance genes and can therefore be cultured in media ...

...is its simplicity and versatility. CRISPR/Cas9 consists of two components: the non-specific endonuclease...libraries (referred to simply as “CRISPR libraries”) consist of a heterogeneous population of gRNA-containing...drug screens to identify genes that confer drug-resistance. Mutant cells are treated with a drug of interest...and gRNA distribution is analyzed in the drug-resistant population compared to a non-treated control group...enriched” correspond to genes that confer drug resistance when mutated. Findings from this type of experiment...shed light on the mechanism by which cells gain resistance to drugs and can identify future therapeutic ...

... to the 5’ end of the reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride...modification, from insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer...are due to incorrect primer sequences! Primers consist of two regions: the 3’ end binds template DNA for...unamplifiable’ Check your amplification by gel electrophoresis. In our experience, good amplification after...rapid method for recombination and site-specific mutagenesis by placing homologous ends on DNA using polymerase...

...incompatibility with antibiotic-resistance plasmids, scientists have expelled antibiotic resistant plasmids from Enterobacteriaceae...partitioning system in the bacteria, but generally it consists of a centromere-like region on the plasmid, a ...plasmid and pair the plasmids together (akin to sister chromatids coming together in eukaryotic cells)...plasmid incompatibility to combat antimicrobial resistance. Using novel plasmids based partly on plasmid...2017) Plasmid interference for curing antibiotic resistance plasmids in vivo. PLOS ONE 12:e0172913 . https...

...Agarose Gel Electrophoresis Inoculating Liquid Bacterial Cultures Restriction Digest Analysis We also...fastest shipping. However, there were issues that persisted despite our best efforts. With a little over half...having some materials available to order, yet not consistently accessible, wasn’t an option. So we identified...γ-Retrovirus Guide Protocols Agarose Gel Electrophoresis Bacterial Transformation How to Design a Primer...Verify Your Plasmid Using a Restriction Digest Analysis Videos What is a Plasmid? How to Design...

...method for transient gene expression analysis in seedlings of Arabidopsis and other plant species." Plant ...   C. roseus is not a model organism such as Arabidopsis or tobacco. Studying gene function in C. roseus...method for overexpressing genes In plants like Arabidopsis and tobacco, Agrobacterium infiltration into ...expression assay by agroinfiltration of leaves." Arabidopsis Protocols. Humana Press, 2006. 225-229. PubMed...of our plant biology blog posts Find tips for Arabidopsis transformation Read our Plasmids 101 blog posts...

...Efficient and Stable Genome integration Insertional mutagenesis, transgene reactivation, residual expression ...Efficient and Stable Genome integration Insertional mutagenesis, residual expression piggyBac Excisible Medium...generating methods due to the risk of insertional mutagenesis. Random integration also creates heterogeneous...Finally, retroviruses are prone to insertional mutagenesis and random integration creates variation between...because they are less likely to cause insertional mutagenesis, adenoviral vectors are considered a safer way...These methods eliminate the risk of insertional mutagenesis, the presence of a genetic scar, and incomplete...replication of the plasmid in mammalian cells and assists in tethering the vector to the cell’s chromosomes...

...the DNA during DNA synthesis and then diluted with subsequent rounds of DNA synthesis (cell divisions)....it comes to labeling cells for flow cytometric analysis, the most common method is a cell surface label... and gate on the positive events for your flow analysis. However, certain downstream applications, like... even at room temperature, allowing for a flow analysis to be shifted to a later day. Note that intracellular... cells can still be easily gated on during the analysis. Conclusion Congratulations! You made it through...

...ddNTP to a synthesis reaction along with all four regular nucleotides results in the synthesis of DNA strands...run on a gel, and transferred to a membrane for analysis through hybridization of a radioactively or fluorescently...blotting, in conjunction with restriction digest analysis, is used to identify the presence of the mutated...sequences among DNA fragments separated by gel electrophoresis." Journal of molecular biology98.3 (1975): ...copy number changes." Protocols for Nucleic Acid Analysis by Nonradioactive Probes (2007): 205-226. PubMed...

...fused to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables selection...requested plasmid, SpCas9 is still going strong as the basis for many genome editing experiments. So, without...ribosome to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This is the ..., which had a point mutation in the puromycin resistance gene that made it less effective in certain cell...

... One consists of NLuc fused to enhanced green fluorescent protein (GpNluc) and the other consists of NLuc...transfer (FRET) probes. These probes generally consist of a responsive protein (e.g. calcium binding protein...now specializes in supporting Promega cellular analysis products at Promega.     References 1. Hall, M.P...optical imaging and flow cytometry for monitoring tumorigenesis. Cancer Res. 75, 5023-33. PubMed PMID: 26424696... (2015) NanoBRET-A novel BRET platform for the analysis of protein-protein interactions. ACS Chem. Biol...

...Cells on the membranes will then be exposed to a lysis buffer to release recombinant proteins directly ...but overnight at 30°C is a good start. Prepare a lysis solution containing your favourite bacterial protein... with thick filter paper and saturate each with lysis solution (~3 ml each), pouring off any excess. When... membranes off the plates and place each onto a lysis solution-soaked filter paper (colony-side up) and...to several freeze/thaw cycles to facilitate cell lysis: alternating between 5 min at -20°C (or -80°C) followed.... Weigele, Peter, and Elisabeth A. Raleigh. "Biosynthesis and Function of Modified Bases in Bacteria and...

... in bacteria, transposons can move antibiotic resistance genes between plasmids or from plasmids to the... how they can be used in the lab. Transposon mutagenesis screens Transposons, by their nature, are mutagenic... elements, which makes them a great tool for mutagenesis screens that detect loss-of-function or gain-...to consider when selecting a transposon for a mutagenesis screen. For example, piggyBac and Tol2 are best...Insertion of Transposable Elements by Guide RNA-Assisted Targeting) system. INTEGRATE is based on a naturally...of which has several different applications (mutagenesis screens, transgenic animals, gene transfer). ...

...In this method, substrate pools were prepared consisting of one target (template) strand and four upstream...identifiable and quantifiable by capillary electrophoresis (CE) (see below). This method allowed screening...gene PCR-ligase detection reaction-capillary electrophoresis assay." Journal of clinical microbiology 45.6...mixed population of excess wild‐type DNA." Electrophoresis 34.9-10 (2013): 1415-1422. PubMed PMID: 23463388...reaction-fluorescent microsphere assay for characterization of resistance-mediating polymorphisms in African samples of...Greenough, Lucia, et al. "Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate...