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We narrowed to 731 results for: Sis;

Showing: 201 - 240 of 731 results

  1. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...complementary to the dsRNA (Wilson and Doudna, 2013). RNAi consists of two non-exclusive gene silencing pathways: ...different Dicer-like enzymes are involved in the biogenesis of different small RNAs (Figure 1). These siRNAs...amiRNAs based upon miRNA precursors of rice and Arabidopsis species, respectively. Short hairpin RNA (shRNA...plants, such as in the model flowering plant Arabidopsis thaliana (Li et al., 2013; Zhang et al., 2018...plants outside of limited, single cell protoplast analysis. Instead, microscopic plants must be visually ...

  2. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...guest blogger, Kaustubh Kishor Jadhav, a Research Assistant at MGMs Institute of Biosciences and Technology.... The absence of a rigid cell wall makes them resistant to antibiotics and antibacterial drugs like penicillin...of antibiotics can allow mycoplasma to develop resistance to the antibiotics. Antibiotics must only be ...prolonged use of a particular antibiotic can induce resistance for other bacteria apart from mycoplasma. Always...Jadhav! Kaustubh Kishor Jadhav, MSc is a Research Assistant at the MGMs Institute of Biosciences and Technology...

  3. Antibodies 101: Validation

    Type
    Blog Post
    ...antibodies as major culprits in the reproducibility crisis of biomedical research (Baker, 2015; Bradbury &...approach. Antibody-independent methods may include analysis of transcriptomic or proteomic data from several...protein interactions could be a factor in your analysis. Example: You are planning an RNA-immunoprecipitation...and send the captured proteins off for mass spec analysis (above). When the results come back it shows that...here!  References Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature 521:274–276...

  4. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ... the cause of leaky expression. To test their hypothesis, they randomly scrambled one pair of lox sites...packaging even begins. This spontaneous recombination persists even in recombinase deficient E. coli and under...References Aschauer DF, Kreuz S, Rumpel S (2013) Analysis of Transduction Efficiency, Tropism and Axonal...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...

  5. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...replace a gene of interest with an antibiotic resistance cassette.   Substrate DNA Design and Generation... selectable DNA fragments, such as antibiotic resistance genes, as well as non-selectable DNA fragments...red phage, λTetR, that carries the tetracycline resistance gene and the lambda red repressor cI857.  Once...confirmation of recombinant clones If a antibiotic resistance gene has been inserted, recombinants can first...first be selected via antibiotic resistance, but all clones should be further tested to confirm the presence...

  6. How Exercise Purges Stress and Keeps Depression at Bay

    Type
    Blog Post
    ...extra PGC-1a1 only in their skeletal muscle show resistance to stressful circumstances – including unpredictable...Activation of the PGC-1a1 Pathway Gene expression analysis of the animals’ skeletal muscles led to another...mck-PGC-1a1 mice, and those who exercise, show resistance to depression. For the researchers led by Dr...t what they’d expected. "Our initial research hypothesis was that trained muscle would produce a substance...

  7. Typing CRISPR Systems

    Type
    Blog Post
    ... sequence similarity, phylogenetic analysis, neighborhood analysis of surrounding genes, and experimental...sequence divergence that requires a multi-pronged analysis to sort effectively. The current classification...are merely linked by a similar function. After analysis, CRISPR classifications were placed into a fairly... of factors, including your goal, experience, hypothesis, and even the time and money available. The breadth...

  8. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...FP-chimeric protein may be at any one of many stages of synthesis and degradation. For this reason it is virtually...PA-FPs by performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry...negative” photoswitching. In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't...is blocked during the Bax activation phase of apoptosis." J Cell Biol 164.4 (2004): 493-499. PubMed PMID...PMC1635685. 12.Paez-Segala, Maria G., et al. "Fixation-resistant photoactivatable fluorescent proteins for CLEM...

  9. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ...higher volume can affect the lysis buffer ratio and result in incomplete lysis. You may be able to scale ..., which can happen if you leave the sample in a lysis buffer for too long or if you get a little overzealous...scale the amount of buffers used to ensure proper lysis, but you will still face column capacity limitations...troubleshooting. If you need a high level of consistency between lots, then reviewing their quality assurance...

  10. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ... more functional benefits to the host such as resistance to antibiotics, degradative functions, and/or...advantageous for survival, such as an antibiotic resistance gene. In contrast, plasmids utilized in the ...recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing... obtained. Selectable Marker The antibiotic resistance gene allows for selection in bacteria. However...

  11. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...Primer design. Plasmid mapping. DNA sequence analysis. We all have our favorite tools for tackling these...plasmid Editor): A free, donation-based plasmid analysis tool including editing, annotating, creating maps...pET21a-BirA.   DNA / protein database search and analysis tools Addgene's analyze sequence tool: Annotates...Assembly  OligoAnalyzer Tool from IDT: For hairpin analysis, Tm and primer dimer estimation, and other primer...

  12. Tips for Improving Your Next Manuscript

    Type
    Blog Post
    ...multiple high-profile publications. My classmates consisted of graduate students, post-grads, and early career...topic by the moderators. The assignments, which consisted of reading that class’s section of a manuscript...Four ways to improve your next manuscript 1. Be consistent with your figures The figures of a manuscript...and they should also be easily understood and consistent in design and layout. 2. Be concise in your writing...

  13. Oh, The Places You Can Go: Careers in Science Communication - Curriculum Development

    Type
    Blog Post
    ...kinematics in physics, to developing a lab about mitosis and meiosis in biology. Lots of it is researching the... to students, like how to pour a gel [for] electrophoresis or doing a dissection video. I did that in ...themselves. “You open up the box and there’s a gel electrophoresis tray in there. Or the dissection specimens....

  14. Safe Port in a Storm...How Addgene is Weathering the Pandemic

    Type
    Blog Post
    ...earlier than most companies. Since the start of this crisis, we’ve been very responsive to the mood of the ...unknown amounts of time, we were facing a financial crisis. As a nonprofit organization, Addgene charges enough...figure out a way to maintain our services during the crisis and maintain our workforce for when we need to ...plasmids and other reagents every day, and this crisis has highlighted the importance of access to quality...

  15. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    ...learn about trends in synthetic biology and DNA synthesis. They published their results in a paper in Nature...biology, in particular, the ability to do DNA synthesis, is evolving so fast that it is a really nice ... in GC content between many organisms and DNA synthesis companies often work within a narrow range for...Philipp Pfingstag, and Neil C. Thompson. "Gene synthesis allows biologists to source genes from farther...

  16. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...The researchers identified MerMAIDS from their analysis of different publicly available metagenomic data... study demonstrates the power of combining the analysis of publicly available metagenomic datasets with...fused dCas9 to a truncated CIB1 protein from Arabidopsis (CIBN), which they call the LADL Anchor. Using... iterations is the addition of CRY2, another Arabidoposis protein that heterodimerizes with CIBN upon ...

  17. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ... lost a specific gene survive (ex: antibiotic resistance marker). In a previous blog post, we covered ...plasmid backbone, as it often relies on antibiotic resistance. But what about identifying clones that contain...Lynne Postovit’s lab. It contains an ampicillin resistance marker in the backbone. A selection on agar containing...visualize this process, or visit our restriction analysis blog post. Colony PCR Colony PCR can detect...

  18. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    ...mechanism by which bacteria have gained antibiotic resistance and virulence. Scientists have studied how HGT... replication-incompetent phage particles with assistance from a ‘helper’ phage prior to transduction. ...can infer genetic advantages such as antibiotic resistance. Bacterial conjugation as explained by...Bacterial Conjugation. In An Introduction to Genetic Analysis. 7th edition (2000). Meibom, Karin L., et al. ...

  19. AAV Q&A with Tim Miles

    Type
    Blog Post
    ...dosing, we advise that users keep titering method consistent across samples within a project. Targeting Your... on vector dose. The color diversity in Vector Assisted Spectral Tracing (VAST) experiments using PHP....best practices’ for model organisms.    Q11: How consistent is expression is when injecting CAP-B10 or CAP-B22...linked to epigenetic changes, such as multiple sclerosis?  A: Basic science and preclinical researchers...

  20. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...of common plasmid features such as antibiotic resistance genes, selectable markers, promoters, origins...plasmid sequence For more immediate, high-level analysis, you can click directly on the “Analyze Sequence...to additional tooltip information and sequence analysis tabs including: Maps This is the first tab...For example, if you hover over the ampicillin resistance gene, you’ll find the full name of the gene, ...

  21. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...expression profiling, systematic tumorigenesis assays, drug epistasis assays, and morphological profiling...huge challenges preventing us from testing this hypothesis: There was no large scale, broadly available... corresponding to cancer variants derived from analysis of lung adenocarcinoma genomes (Lung Cancer Mutant...

  22. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the...basepairs or small indels that occur during DNA synthesis, and protomoting or repressing protein expression...cleave DNA. DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired...Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org...

  23. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...will catch you up on the core replication and resistance features of yeast vectors and explain how they... and diploid state which is useful for genetic analysis, as well as an efficient mechanism of homologous... due to high rates of spontaneously occurring resistant mutants and the insensitivity of yeast strains...auxotrophic markers. The MX series of antibiotic resistance cassettes is most commonly employed, with the...

  24. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...Protein turnover is the combined effect of protein synthesis and degradation. These processes control the abundance...possible to visualize the localization of protein synthesis. Alternatively, fluorescent protein tags can be...the system. Another method for studying protein synthesis in situ is the knock-in of fluorescent tags into...Nathans, Daniel. "Puromycin inhibition of protein synthesis: incorporation of puromycin into peptide chains...

  25. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...that is resistant to the toxic effects of CcdB. Another commercially available, CcdB-resistant strain ... investigator’s gene of interest.   CcdB resistant E. coli strains complete the system But ccdB alone...contains the F plasmid (F’ strains) will also be resistant to CcdB, as the native ccdA will be present. Most...

  26. Plasmids 101: Cre-lox

    Type
    Blog Post
    ... of replication, protein tags, and antibiotic resistance markers (just to name a few). In this edition...site-specific recombination events. The system consists of two components derived from the P1 bacteriophage...sites are 34-base-pair long recognition sequences consisting of two 13-bp long palindromic repeats separated...targeting, targeted clones are selected for using a resistance marker; however, it is often desirable to remove...

  27. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    ...gotten myself.” Perkins noted that there is a more emphasis today about non-academia and industry jobs. Since...her first job at Addgene as an Order Processing Assistant. After a few months, she left school when she ...mean you’ve failed. “I got a job as a research assistant in a lab at the university that I went to. Making.... She was working as a research and technical assistant, when her life changed––she had a daughter. Needing...

  28. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ...Kleckner. "Recombination, pairing, and synapsis of homologs during meiosis." Cold Spring Harbor perspectives...Special breeding cases Whether your mouse colony consists of genetically engineered mice with a single mutation...chromosome. Homologous chromosomes recombine during meiosis, creating gametes that contain a mix of genetic...

  29. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ... 1000000097) The Winslow lab barcoding library consists of 96 retroviral plasmids (based on the Murine...high-throughput sequencing to perform multiplexed analysis of cells pre-treated with a compound of interest...Jacks. It carries CRE recombinase and a puromycin resistance cassette. No time to produce virus? Get the ...mammalian cells. The MuLE system kit from the Frew lab consists of 91 building block Entry vectors and 5 lentiviral...

  30. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...makes them ideal candidates for applications in oncolysis and vaccination!) In the 65 years since its discovery...virion is a non-enveloped particle of ~100 nm, consisting of a capsid surrounding an inner core which contains...adenovirus: the virion is ~ 100nm in size, and consists of an icosahedral capsid made of 3 types of proteins... for creating adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the ...

  31. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...generating huge amounts of data that could provide the basis for student projects in introductory labs. Genome...evolution of the genes involved in methionine synthesis (5). The budding yeast, Saccharomyces. cerevisiae...deletion strains is available (6). Methionine synthesis in S. cerevisiae (Figure 1) occurs through a well-characterized...cerevisiae genome by gene deletion and parallel analysis. Science 285, 901-906. (1999). PubMed PMID: 10436161...

  32. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...the mRNA molecule, forming a ribosome complex consisting of the translated protein (sybody), ribosome,...constructed, they are used to produce a phage library consisting of phages that display the candidate sybodies...tag-free proteins for further characterization and analysis. In a rapid response to the COVID-19 pandemic,...Schepens B, Saelens X, McLellan JS (2020) Structural Basis for Potent Neutralization of Betacoronaviruses by...

  33. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...customization with strain-specific promoters, antibiotic resistance selection markers, gRNAs, and dCas9 proteins....Mobile-CRISPRi can be used to identify antibiotic resistance genes. Genes that are targets of antibiotics ...essential genes such as those involved in antibiotic resistance. Are you ready to move Mobile-CRISPRi into your...30675034. Peters, Jason M., et al. "Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi." Nature...

  34. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...plants that inherited an edited gene to confer resistance to the herbicide chlorsulfuron (Homo line 2) ... to drive prime editing in both tomatoes and Arabidopsis, suggesting the improvements may be broadly applicable...enables heritable desired edits in tomato and Arabidopsis. Nature Plants, 10(10), 1502–1513. doi: https...the formation of Gα-GTP by BRET. The biosensor consists of a YFP-tagged G protein and a NanoLuc® (Nluc...

  35. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...Patrizia, et al. "Ostreopsis cf. ovata induces cytoskeletal disorganization, apoptosis, and gene expression...Additional passages will allow your cells to achieve a consistent growth rate. For every minute spent organizing... 1. Yin, R., et al. "Gene expression profiling analysis of bisphenol A-induced perturbation in biological...

  36. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ...plants, and agricultural crops in particular, resistant to disease. They also devise biosensors designed... developed mostly with model plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana is ...Universities sign up and in the first year there is an analysis of women in faculty, how they are doing. They ...Plant SynBio Reading: Quick, Versatile Plant Transgenesis with GreenGate Plasmids  ...

  37. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...the alignment at a more distal amino acid. We consistently run our blastx searches with the “Low complexity...recommendation is not infallible, we have found it saves analysis time to remove this default setting. Timesaving...Depending on your sequencing result and desired analysis, BLAST may not always be your optimal choice. ...plasmid maps. Resources on Addgene.org Sequence Analysis of your Addgene Plasmid Addgene Plasmid Guide ...

  38. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...specific genomic region using the CRISPR system, consisting of dCas9 and targeting gRNA, to identify molecular...vitro" form. In this form, the CRISPR complex, consisting of a recombinant dCas9 protein and synthetic ...Yuno, and Hodaka Fujii. "An enChIP system for the analysis of bacterial genome functions." BMC research notes...expressing the 3x FLAG‐dC as9 protein for enCh IP analysis." Genes to cells 23.4 (2018): 318-325. PubMed ...

  39. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ... rare genetic eye disease, Leber’s congenital amaurosis. This is the first of what I, scientists, doctors...disease more prevalent or life-threatening (cystic fibrosis, hemophilia, muscular dystrophy)? AAV gene therapy...childhood-onset form of blindness name Leber’s Congenital Amaurosis (LCA). Shortly after the cloning of the RPE65 ... time and place. But it is the hard work and persistence of a dedicated team of scientists led by Dr. ...

  40. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...primarily European reference genome could affect their analysis. Their work revealed that cell lines from individuals...information, Misek notes that in theory, if the DepMap analysis had an equal representation of every ancestry ...descent.Correcting the bias  While their thorough analysis uncovered some concerning biases within widely... team at Broad, they were able to change their analysis to use a corrected genome map. Now, any data that...
Showing: 201 - 240 of 731 results