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We narrowed to 608 results for: Sis

Showing: 81 - 120 of 608 results

  1. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    Published
    March 8, 2021, 2:41 p.m.
    ...should be consistent across all samples on the gel and should be observed as a band of consistent size and... Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is the first step of a western. ...separate by size. Smaller proteins encounter less resistance in the acrylamide matrix and migrate faster through...percentage of acrylamide to increase migration resistance and improve separation. For large proteins, reduce...it’s time to analyze your protein of interest. Analysis typically begins by confirming that the protein...loading or incomplete protein transfer causes inconsistent levels of the loading control. In these cases...

  2. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    Published
    Aug. 8, 2019, 1:19 p.m.
    ...data analysis/visualization using R/Shiny. She is passionate about open science, BIG data analysis and ...interpretations. A good peer-reviewer will help spot inconsistencies in the work, missing information that would... specific repositories (Addgene, WormBase, Arabidopsis.info). These repositories will ensure that the ...induced responses in Root System Architecture of Arabidopsis HapMap population in Amsterdam. Currently, she...salt-induced changes in root-to-shoot ratio in Arabidopsis, (2) expression patterns in plants with enhanced...

  3. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    Published
    Dec. 13, 2018, 1:14 p.m.
    ...fact, due to impaired endocytosis. In parallel assays designed to test the endocytosis of a fluorescent dye...strategies or developing novel methods for data analysis, our team’s goal is to provide scientists with...cell membrane and are taken into the cell via endocytosis. In our study, we aimed to prevent the uptake...packaging cells by treating the cells with the endocytosis inhibitor sucrose. Previous work by Stoneham ...Hollinshead, and Amin Hajitou. "Clathrin-mediated endocytosis and subsequent endo-lysosomal trafficking of ...

  4. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...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...

  5. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    Published
    Dec. 18, 2014, 4:06 p.m.
    ...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...

  6. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ...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...

  7. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    Published
    Sept. 8, 2020, 2 p.m.
    ...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...

  8. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...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...

  9. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...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 ...

  10. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...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 ...

  11. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...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 ...

  12. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ... 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...

  13. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    Published
    March 31, 2020, 1:15 p.m.
    ...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...

  14. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ...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...

  15. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ...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...

  16. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...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...

  17. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...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...

  18. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ...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...

  19. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ... 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...

  20. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...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...

  21. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    Published
    Jan. 11, 2017, 8:47 p.m.
    ...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...

  22. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ... 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). ...

  23. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ...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...

  24. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    Published
    May 18, 2023, 1:15 p.m.
    ...cytotoxicity (ADCC) c) antibody-dependent cellular phagocytosis (ADCP). Figure adapted from van Erp, et al.,... degranulation.  Antibody-dependent cellular phagocytosis (ADCP) So far, our antibodies have induced “...for is eating. In antibody-dependent cellular phagocytosis (ADCP), Fcγ receptors on macrophages bind IgG...antibodies are bound to in a process known as phagocytosis.  Other functions We’re still learning about...et al. Promoting antibody-dependent cellular phagocytosis for effective macrophage-based cancer immunotherapy.Sci...

  25. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    Published
    Dec. 21, 2017, 2:06 p.m.
    ...researchers manipulating DNA sequences on a daily basis, many found these tools inadequate for planning,... plasmids contain features such as antibiotic resistance markers and replication origins, but there is...feature. With the increasing popularity of gene synthesis, many researchers now use codon-optimized versions...control elements in commercial plasmids were inconsistent and sometimes clearly wrong. In an effort to...Additional Resources on Addgene.org Example plasmid analysis powered by SnapGene Browse All Plasmids ...

  26. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ...usefulness of CPPs must be evaluated on a case-by-case basis. While setting up their CPP system to study protein...coined the term iTOP (induced transduction by osmocytosis and propanebetaine) to describe this method of...studies showed that iTOP functions through macropinocytosis. D’Astolfo et al. tested iTOP in a series...primary (hESCs) and cultured cells showed up to 70% resistance to diphtheria toxin, indicating a 70% frequency...macromolecules into cultured mammalian cells by osmotic lysis of pinocytic vesicles. Okada CY, Rechsteiner M. ...

  27. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    Published
    July 21, 2017, 12:48 p.m.
    ...hedgehog under a new promoter each time a new hypothesis is tested. Controlling multiple genes with Gal4...Drosophila : A Fly Geneticist’s Swiss Army Knife, Genesis 34, 1–15 (2002).PubMed PMID: 12324939. 3. S. E....Davis, Gene expression systems in Drosophila: A synthesis of time and space, Trends Genet. 20, 384–391 (...Desplan, Power tools for gene expression and clonal analysis in Drosophila, 9, 47–55 (2012). PubMed PMID: 22205518...transgene expression, lineage tracing, and mosaic analysis., Cell 141, 536–48 (2010). PubMed PMID: 20434990...

  28. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...for a repair template (sister chromatid or homologue) or extensive DNA synthesis. NHEJ also finishes repair...Cheng A, Hasty P, Stark JM (2009) Limiting the Persistence of a Chromosome Break Diminishes Its Mutagenic...Ding X-Y, Guo X-P, Jin Q, Sun Y-C (2020) A CRISPR-Assisted Nonhomologous End-Joining Strategy for Efficient...Efficient Genome Editing in Mycobacterium tuberculosis. mBio 11: . https://doi.org/10.1128/mbio.02364-19 More...

  29. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...stranded and its ribose groups are susceptible to hydrolysis and heat degradation. Furthermore, RNases, or...without something to block RNase activity upon cell lysis, RNA is degraded. To effectively isolate intact ...RNases had a chance to break down RNA upon cell lysis.  In the late 1970’s, Chirgwin and colleagues showed...your samples cold until homogenization and cell lysis. This can be challenging depending on your laboratory...ways to “stabilize” RNA within tissues before cell lysis. These products, namely RNAlater®  from Thermo and...

  30. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    Published
    April 23, 2018, 2 p.m.
    ...antibody. TRIM21, an E3 ubiquitin ligase, is the basis of TRIM-away. In normal physiology, TRIM21 recruits... to their small size (<250 nm), and studying endocytosis is further complicated by the reliance on pH-...addition, the approach allows minute-by-minute analysis of the kinetics of chromatin regulation and subsequent... reversibility allows precise and quantitative analysis of epigenetic memory at nearly any locus. The ...genes of interest. These transcription factors consist of the GAL4 or ZIF268 DNA binding domains fused...

  31. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...no small task, however, at a repository that consistently receives around 200 new DNA samples every week...inserted gene/functional mutations, antibiotic resistance, restriction sites used during cloning, and most...our other blog posts for more tips on sequence analysis, plasmid verification, and using NCBI BLAST. Tips...and read the submission instructions. As for the analysis of a large number of sequencing reactions, you... be done through the align tab of our sequence analysis web-tool. Alternatively, you can use DNA-editing...

  32. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    Published
    Jan. 21, 2015, 1:30 p.m.
    ... Your Thesis” online course.  To learn more about her site please visit www.FinishYourThesis.com.   Additional...post was contributed by Dora Farkas of www.FinishYourThesis.com and is one of many posts in our career...have trouble making decisions. In my work as a thesis coach for graduate students, I heard about supervisors...Ph.D. from MIT and is the Founder of Finish Your Thesis, an online community that has supported thousands...

  33. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ...through directed evolution Hu et al. used phage-assisted evolution to generate and select for Cas9 mutants...mutants with PAM flexibility. In this system, the mutagenesis plasmid (MP6) permits quick, large-scale mutation...and subjected them to another 72 hours of phage-assisted evolution, generating xCas9 2.0-2.6. This second...where cleavage leads to the loss of spectinomycin resistance. xCas9 3.7 showed the broadest PAM range, recognizing... PAM for SpCas9. Hu et al. performed GUIDE-seq analysis to characterize genome-wide off-targets, since...

  34. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    Published
    Sept. 24, 2024, 1:15 p.m.
    ...eliminate sources of interference, but the result persisted. (Even better, multiple other labs have reproduced... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...as light-induced conformational changes are the basis of many optogenetic tools. And, since MagLOV’s parent...Magnetoluminescence from a Spatially Confined Persistent Diradical Emitter. Journal of the American Chemical...

  35. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ... DNA and look for potential contaminants. The analysis is a 2-step process using Geneious software: First...open-access Python scripts that can perform a deeper analysis of the data. Our serotype detection software confirms...Adeno-Associated Virus Production and Their in Vivo Persistence after Vector Delivery. Molecular Therapy 12:744...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...

  36. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...suppressors, and their protein products. Proto-oncogenes consist of a group of genes/proteins that, when expressed...awry. The process of programmed cell death, or apoptosis, is one such mechanism. When a cell or its genetic...project continues to evolve rapidly and, with assistance from the research community, will soon expand...pathways with potential to confer anticancer drug resistance." Science signaling 7.357 (2014): ra121. PubMed...Resources on the Addgene Blog Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer...

  37. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...homology, allowing the strands to anneal. DNA synthesis then fills in the gaps. The end result is a deletion...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus...into the FBL locus. Upon sequencing of puromycin resistant clones, they found that 80% and 50% of clones ...References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with...and Ling Lu. (2016). “Highly efficient CRISPR mutagenesis by microhomology-mediated end joining in Aspergillus...

  38. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...namely that they can cause cancer via insertional mutagenesis (which HIV does not do). Furthermore, molecular... thanks to Addgenie Rebecca Hutcheson for her assistance with lentiviral replication.  Resources and ...1974). Infectious rous sarcoma virus and reticuloendotheliosis virus DNAs. Journal of Virology, 14(5), ...polymerase of Rous sarcoma virus: Initiation of synthesis with 70 S viral RNA as template. Journal of Molecular...Full-Length Human Immunodeficiency Virus Type 1 DNA Synthesis. Journal of Virology, 82(5), 2376. https://doi.org...

  39. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ...recombination.    Designing donors for HDR The basis of the HDR pathway relies on repair from a template...molecule, which is usually an endogenously available sister chromatid, though exogenous DNA can be used as ...kb), a dsDNA donor will need to be used due to synthesis limits of oligonucleotides. These donors traditionally...predominantly in S and G2 phase of the cell cycle, when a sister chromatid is likely to exist as a repair template...long. Prolonged cell cycle arrest can lead to apoptosis in as little as 10 hours, so read up on the timing...

  40. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...human CRISPRko sgRNA library The Brunello library consists of 77,441 sgRNAs with an average of 4 sgRNAs per...depletion of essential genes. In the subsampling analysis of the sgRNAs, Brunello even with only a single... a comparison study screening for vemurafenib resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic...Mediator) uses an improved activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA...reading frame (ORF) overexpression libraries for resistance to MEK inhibition. Although both Calabrese and...
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