...mechanisms of the host. The cause, manifestation, diagnosis, recurrence, and treatment of cancer have been... process analogous to the development of drug resistance in bacteria, the tumor cells that escape the ...is a region of incessant chemical activity, and consists of other cell types, especially immune cells and... promotes the production of VEGF to increase angiogenesis around the tumor. This in turn regulates the... tumor cells spreading to other organs during metastasis. Evading Recognition Tumor cells over-express...damage healthy cells, and often cannot eradicate metastasis or eliminate the probability of a relapse. Chemical...the immune cells that aid tumor formation and metastasis, block the cell-surface proteins that help escape...

...including but not limited to cell growth and drug resistance. Recently, the Wu Lab at Texas Tech developed...Virus (WNV) induced cell death. The Wu lab library consists of 77,406 individual gRNAs targeting a total of...approved drugs against the entire kit; a parallel analysis that successfully identified new, highly specific...alone involves much literature searching, DNA synthesis, and plasmid assembly. Luckily, researchers from...Moduclar Cloning (CIDAR MoClo) Parts Kit. This Kit consists of a variety of plasmids containing many different...processes such as differentiation, proliferation, and apoptosis. In humans, mutations that permanently activate...plasmids to the scientific community. This collection consists of plasmids containing genes from a crowd-sourced...

...for the analysis. (C) A more liberal gate allows more cells to be considered for the analysis. Depending...duplicate events, and the Live/Dead gate to focus your analysis on what’s biologically active. After that, the... of tissue processing prior to running my flow analysis, I can however clearly identify my lymphocyte ...good to ask around for advice before running your analysis. In this case, I and other lab members had worked...acquisition voltage (not shown) used in my flow analysis I can spot my lymphocyte population distinct from...beginning should not have a great impact on your analysis, and I often choose a looser gate here. I am not...worried about including irrelevant events in my analysis, as these will get excluded through further, more...

...plasmid contains a cassette that will make cells resistant to a toxin), whereas negative selection would ...markers are listed below: Name Gene Conferring Resistance Cell Types* Mode of Action** Working Concentation...CHO, 293HEK, Jurkat T cells Blocks polypeptide synthesis at 80S; inhibits chain elongation 100-800 ug/mL...NIH3T3, CHO, Jurkat T cells Blocks polypeptide synthesis at 80S; inhibits chain elongation. 50-500 ug/mL...HeLa, 293HEK, Jurkat T cells Inhibits protein synthesis; premature chain termination 1-10 ug/mL Zeocin...reagent). Cells should die within 3-5 days and resistant colonies appear in about 10-14 days depending ...confusing since the neo/kan gene confers G418 resistance; however, like gentamicin, neomycin is typically...

...data for a hypothesis and presents only the results that align with the original hypothesis. Other practices...steer clear of saying we have a reproducibility “crisis” on our hands and instead provide a brief conceptual...in-depth information about reagents, protocols, analysis tools, and more. Being that this is the Addgene...larger and larger datasets and use computer based analysis techniques, an important component of reproducibility...researchers to share code they’ve used for data analysis while other organizations like Dryad make it easier... stories with links of causation from initial hypothesis through validation are easier to understand. ...publication record will then show that the tested hypothesis is correct (a great success story) even though...

...integrating sex and gender analysis into research. Why are sex and gender analysis important? Doing research...and gender analysis and 2) provides case studies with concrete examples of how gender analysis leads to ...harnesses the creative power of sex and gender analysis for innovation and discovery. Tackling sex and... the potential to save lives and money. A 2014 analysis of the US Women’s Health Initiative Hormone Therapy.... While most of the results were positive, the analysis did find 263,000 more osteoporotic fractures. ... learn sophisticated methods of sex and gender analysis. Universities need to incorporate these methods...harnessing the creative power of sex and gender analysis for innovation and discovery. She is a member ...

...antibodies are, they are in a midst of a crisis. The “antibody crisis” came to light as antibodies became ...2015 review analyzing and explaining the antibody crisis states that up to 50% of all commercially available...Plückthun, 2015). To avoid being caught in the antibody crisis, one must corral and corner the goldilocks antibodies...scientists to confidently choose reliable and consistent antibodies for specific markers. The Screening...determine the antibody’s reliability through their own analysis of the background noise, distinct imaged pattern...bring us one step closer to resolving the antibody crisis. The mAb3D Atlas is a public service to the science...please contact us here! Ava Shipman is a research assistant in lab of Zhuhao Wu at the Icahn School of Medicine...

...functional antibiotic resistance. Colonies with an insert in pUCXKT will therefore have resistance to both kanamycin... the lambda phage integrase system which is the basis for Gateway cloning. The combination of all 11 att...activator protein to drive transcription. The anchor consists of dCas9 fused to CIB1 and is targeted to specific...readily modified to use different antibiotic resistances, restriction enzymes and plasmid backbones. pUCXKT...nonfunctional truncated version of the kanamycin resistance gene (missing the first two codons). Two stop... promoter to prevent leaky expression of this resistance. Unlike previous systems, pUCXKT does not result...containing the missing codons from the antibiotic resistance cassette, a ribosome binding site, SacI restriction...

...before your enzyme lysis. For cell cultures, enzyme lysis is typically sufficient.  Lysis buffer When choosing...specificity, lower cross-reactivity, and are more consistent lot-to-lot. The highest reproducibility comes...choosing your lysis buffer, consider the subcellular location of your protein. Proteins located in membrane-bound...mitochondria or nucleus, will likely require a harsher lysis buffer than proteins easily available in the cytoplasm...or whole-cell lysate. A harsher, SDS-containing lysis buffer, like RIPA, should be considered if your ...membrane-bound protein. There are many different lysis buffers available commercially, and most common ...use of hazardous chemicals, and are not always consistent. Precast gels, while expensive, reduce protocol...

...chloramphenicol-resistant strains, we have also included versions with gentamicin-resistance (amplified from...disrupt expression of a gene with an antibiotic resistance cassette. This leaves a “scar” in the chromosome...be accomplished by selecting for an antibiotic resistance marker from the vector (abR, Figure 1). The resolution...This vector contains sacB, an chloramphenicol resistance cassette, and the R6K origin of replication to...replicated in the recipient, the only antibiotic resistant colonies isolated are those where the allelic ...crossovers, they are just spontaneous antibiotic resistant clones. But if they are blue or magenta, they ...hospital-associated Gram-negative bacteria become resistant to antibiotics! References Espah Borujeni, Amin...

...treated a tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102,...EPISOME-MEDIATED TRANSFER OF DRUG RESISTANCE IN ENTEROBACTERIACEAE VIII: Six-Drug-Resistance R Factor.” Journal of...other extrachromosomal particles, particularly Resistance or R-factors. Like F-factors, R-factors could... selected for E. coli transformants that were resistant to both. When this proved successful, pSC101 became...Lederberg, Joshua. "Cell Genetics and Hereditary Symbiosis" Physiological Reviews 32.4 (1952) 403-430. Pubmed... PMC314833. Datta, Naomi. “Transmissible Drug Resistance in an Epidemic Strain of Salmonella Typhimurium...

...official thesis lab Undergraduate student Roles and responsibilities can vary "Lab assistants" usually...materials, lab maintenance Administrative assistant: assists PI with admin duties Staff scientist: similar...the lab Official title is typically Professor, Assistant Professor, Associate Professor Sometimes teaches...Graduate student Master's or PhD student Works on a thesis project to earn a degree "Rotation student" - some...usually wash glassware, prep media, etc. "Research assistants" are more involved in the science, experimental...independent project. Lab technician / Research assistant Typically has a Master's or Bachelor's degree..., as well as graduate students (your teaching assistants), about their experiences. Research requires ...

... Open source image analysis software tools can be used for colocalization analysis. Some examples include...of quantitative colocalization analysis methods and useful image analysis software tools. Pitfalls, limitations...simply “looking at them”. Modern cameras and image analysis software are highly sensitive to detecting light..., and customized CellProfiler pipelines. Image analysis software produces quantifiable results and statistics...different experimental setups. A colocalization analysis example is given in Figure 2 (adapted from 4) ...Proteins. Ruusuvuori, Pekka, et al. "Quantitative analysis of dynamic association in live biological fluorescent...

...vector that achieves persistent transgene expression in certain cell types (Tatsis et al., 2007), potentially...expression, and protein translation. Its genome consists of “early” (E) and “late” genes, and the transition...potentially because some of the wild-type viruses can persist as a latent infection in certain tissue reservoirs....1004821. PMID: 26042599. Pereira, H. G. (1972). Persistent infection by adenoviruses. J Clin Pathol Suppl...doi.org/10.1038/s41586-023-05949-1. PMID: 36996871. Tatsis, N., Fitzgerald, J. C., Reyes-Sandoval, A., Harris-McCoy...., & Ertl, H. C. J. (2007). Adenoviral vectors persist in vivo and maintain activated CD8+ T cells: implications...very late stages of infection for efficient cell lysis and release of adenovirus from infected cells. J...

...systems that are widely present in bacteria. They consist of a toxin which can affect a variety of cellular...division”). The encoded toxin-antitoxin system consists of CcdB (toxin) and CcdA (antitoxin) where CcdB...sequences in RNA resulting in a blockage of protein synthesis. The antitoxin MazE, which binds and inhibits ... contains the mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked...) contained within it by selecting for Zeocin resistance. Then, counter-selection with methanol medium...systems Because restriction modification systems consist of two enzymes that have opposing activities, it...the systems just described, the hok/sok systems consist of three components: Host killing (hok), a long-living...

... in every experiment-based investigation of a hypothesis: proper controls! For every experiment, an investigator...ability of the cells to express the protein.   In analysis of the result in Figure 1, an internal control...includes the aforementioned internal control and is consistent with the result in Figure 1 could look like this...associated with the independent variable should be consistent over time and from different preparations of ...replicate control is important, it speaks to the consistency of the reagents and hands performing the experiment...experiment, not the reproducibility or consistency of the observed phenotype associated with the independent...replicates control for the reproducibility and consistency of the observed phenotype associated with the...

...actin and myosin, and enzymes for transmitter synthesis. Retrograde transport is used for transport of...vivo, microinjection, pressure injection or iontophoresis (applying small electric currents via inserted...methods are naturally occurring viruses that infect, persist, and migrate within neurons, and that are also ... Not all measures of transport efficiency are consistent which may be caused by the diversity of techniques...are not fully understood and may be a form of transcytosis where the AAV particle is taken up on one side...cross the blood brain barrier by endothelial transcytosis (Foust et al., 2009; Merkel et al., 2017), and...Aschauer, D.F., Kreuz, S., and Rumpel, S. (2013). Analysis of transduction efficiency, tropism and axonal...

... (2016) Enhanced Rice Blast Resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF Transcription Factor...longer shelf life and the development of disease-resistant crops to increase agricultural yield (Wang et ...including commonly used model organisms like Arabidopsis and Medicago truncatula and several crop species...sequence and ~75 nucleotide scaffold sequence) is consistent between plants and other organisms, but the promoter...GOI) into plant cells. A) The system typically consists of a Ti plasmid, into which the GOI is inserted...2013) Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas...homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew. Nat Biotechnol 32:947–951 ...

...antibiotic resistance cassette and you aren’t introducing an additional antibiotic resistance cassette ...where the inserted DNA includes an antibiotic resistance marker. After transformation, plate the cells...into the plasmid backbone contains a kanamycin resistance cassette, you would plate the cells on kanamycin-containing...cells carrying plasmids that contain a kanamycin resistance cassette will grow. However, often times, the...often used for cloning in yeast since antibiotic resistance spontaneously arises in yeast. For example, Markus...occurring genes for cloning purposes (antibiotic resistance genes, toxin genes, and the sacB gene). By tapping...

...salts. Capillary electrophoresis Quantifying DNA using capillary electrophoresis is similar to quantifying... using gel electrophoresis. Except, it’s smaller. And automated. Unlike gel electrophoresis, you only ... sample concentration for you. Agarose gel electrophoresis Though not the quickest way to quantify DNA...nanofluidic channel and the sample is separated by electrophoresis. Smaller fragments migrate quicker than larger...quantification of a RNA ladder using capillary electrophoresis which shows fluorescence units over time. Smaller...