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Showing: 521 - 560 of 974 results

  1. 5 Great Apps for Lab Life

    Type
    Blog Post
    ...tried to find an app that could help me with calculations. This is how I came across the Gibco Cell Culture...culture and include a counter, lab timer, dilution calculator, and a couple of other nice features. This app...sequence for your enzyme of interest. Lastly, the Tm calculator helps you set up an optimal PCR depending on ...apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme Tool by Promega Material...

  2. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...investigate the effect of regulatory elements — such as promoters, enhancers, or untranslated regions — on gene...Luciferase reporter assays for monitoring gene regulation Since firefly luciferase was first cloned in ...gene expression. The regulatory element is cloned upstream of the luciferase gene, and expression of luciferase...

  3. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ... was the molecule of interest (analyte), and accumulation of ATP activated GFP expression (receptor), ..., then the transducer could be a microscope, plate reader, or any other instrument which measured GFP ...advantages over other mechanisms. For one, many viral platforms, such as AAV or retrovirus, can either permanently...Fluorescent Biosensors for Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers in...

  4. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...of your library cargo to isolate the effects of each variable being manipulated. Thus, the adjustable nature...expression control Researchers are always wanting to manipulate gene expression – turn it up, turn it down, knock...PMC2075523. Bulcha, J.T., et al. Viral vector platforms within the gene therapy landscape. Sig Transduct...

  5. Antibodies 101: ChIP

    Type
    Blog Post
    ...of interest (4) isolated your antibody:protein:DNA complexes and finally (5) isolated the DNA from your...DNA are no longer bound, and the DNA can then be isolated and studied through a variety of downstream methods...downstream analysis via any methods suitable for isolated and purified DNA, such as sequencing or qPCR. ...

  6. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    ... to glance at as you work. Draw your plate maps onto your plates, alongside your physical copy. If you...reminders on a sticky note   Visual Planning  Plate maps, pre-labeling tubes, and even drawing out experimental...example, setting up your pipette tips to match your plate layout can help you track samples as you add them...

  7. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ... in the nucleus. The Roth lab showed that this platform accurately identified 120 known GPCR targets demonstrating..., the receptor fused to APEX2 catalyzes the biotinylation of proteins near the GPCR. These proteins are... GPCRs for chemogenetic studies to probe the relationship between neurons and specific behaviors. Chemogenetics...DREADDs are ideal for chemogenetics as they are relatively insensitive to their endogenous ligand, acetylcholine...

  8. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...multiplexing vectors are larger because they require regulatory sequences to allow for expression of multiple...example of how this works. Make sure to order 5’ phosphorylated oligos or treat with T4 PNK. Delivery options... are easier to use across multiple expression platforms: transfection, lentiviral transduction, and AAV...Multiplex gene editing with CRISPR-Cpf1 is one of the latest developments in CRISPR technologies.  It’s a simple...

  9. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ...participants in skills useful for future mentoring relationships.   If you would like to start a group mentoring...for the time commitment of a formal mentoring relationship or there aren’t enough senior mentors available...ideas on what to do once you are in a mentoring relationship.  Celebrate your successes and support each... come. Are you involved in Peer Mentoring relationships? Comment below with ideas from your own mentoring...

  10. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...The modularity of antibodies means that it is relatively easy to physically divide up the different regions...you to use just a fragment of the antibody in isolation from another portion. For example, Fab fragments...but not the Fc region (more on when that might be later). Find scFv plasmids at Addgene Depending on how...fragments intended for use as primary antibodies are relatively uncommon. Though, if you want to use Fab fragments...

  11. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...mechanism? Where is the control happening? Is the regulation of an exogenously introduced gene or over the...generate a frameshift mutation that will permanently ablate protein expression. If you need the ability to...different amounts of tetracycline. Tetracycline regulation is reversible (when the drug is removed expression... These control systems function at the post-translational level, unlike most of the other mechanisms described...

  12. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ... with post translational modifications. One of the most common of these is 2’-O-methylation, where a methyl...once you’ve chosen a gRNA and modification(s) Congratulations! You’ve picked your perfect guide; now check...Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation. ACS 6, 5, 695-703 (2020). https://pubs.acs.org...

  13. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    ... TAK 1, an enzyme that controls apoptosis and regulates transcription. Without TKA1, cells proliferate...cancer as a broad phenomenon and not only as an isolated disease. While the tumor may seem to take center...microRNA transfer: A mechanism of environmental modulation of hepatocellular cancer cell growth." Hepatology...resources on the Addgene blog Read our cancer related blog posts Learn about our cancer collection pages...

  14. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ... not linearly related to power, the conclusions reached at high power may not translate to real applications...widefield imaging fluorophores are excited with relatively low light levels (for a prolonged time). Other...application (Goedhart et al, 2012). The third issue relates to the environment of the fluorescent protein. ...

  15. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ... that underlie complex regulation in biological systems. The CombiGEM platform provides a means to create.... CombiGEM-CRISPR: The marriage of two simple platforms CRISPR has revolutionized how we decode the genome...Wong et al., 2016). With CombiGEM and CRISPR platforms now being integrated, we look forward to the realization...

  16. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...samples through our lab using barcoded tubes and plates. The critical pieces of information for a given...100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above). We then ... database of over 2000 primers. Two 96-well PCR plates are prepared with the primer stocks for forward...shipped along with their corresponding glycerol plate on dry ice to the sequencing provider. Each sequencing...

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

    Type
    Blog Post
    ...acquired divergent functions due to mutations accumulated over time? Undergraduates can help to answer ...If the phenotype is corrected, students can be relatively confident that the gene from the test species...the S. cerevisiae met deletion strains and then isolate transformed strains on media that lack uracil, ... MET genes to grow.) Students then use replica plating to determine if expression of MET/Met genes from...

  18. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...pluripotency marker expression and response to LPS stimulation, respectively. Primary cell type Gene targeted... a fluorophore or tag. To utilize HDR, a donor template containing the desired sequence flank by homology...Nanoblades which carries a gRNA, Cas9, and a HDR donor template DNA can be generated by incubating Nanoblade particles...facilitated HDR with either a ssDNA or dsDNA donor template. In vivo editing of mouse zygotes and mice with...

  19. Hot Plasmids - October 2020

    Type
    Blog Post
    ...glycosyltransferase genes and other proteins related to cellular glycosylation. Get the library at Addgene CasRx...requires the much less efficient double stranded templates, rather than the more efficient single-stranded...cellular protein. They used CRISPR with a ssODN template to introduce the small wrmScarlet11 which is less...

  20. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...pockets. They allow researchers to identify and isolate very specific biological targets. However, as popular...identical antibodies (See Fig 1.). rAbs are a relatively new technology that takes advantage of DNA sequencing... NOT align with the atlases/literature and is unrelated to the target protein, we categorize the antibody...strong signal. However, exposure time and its relationship to signal depend on what you’re comparing your...

  21. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...donor-acceptor pair and a change in FRET efficiency correlates with a change in the distance and/or orientation...available. Fortunately, several excellent reviews have collated data from various papers to make the decision ...conformation of the probe. More on these biosensors later on). Experiments using intermolecular FRET probes...simultaneously would further increase the amount of correlated information available from a cellular process...

  22. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...embark on or continue their careers in science-related fields. In particular, two recent events have the...note is a ruling in August by the National Labor Relations Board to allow graduate students who work as teaching...university and alterations to the student-university relationship, along with, of course, union dues. Time will...+ adjustments for regional cost of living and inflation) and benefits that were laid out by a 2014 National...

  23. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...typically used (e.g., use 5 mL medium on a 10 cm plate). By reducing the culture volume into which the ...transfection and should give rise to a surviving population of approximately 20–50% of the packaging cells...vector using the harvested viral solution as a PCR template. Regardless of the type of virus you’re working..., and allow you to standardize your experiments later. Stay tuned to the Addgene blog and be sure to check...

  24. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...barely fits into AAV when you add in a gRNA and regulatory elements. To solve this problem, the Zhang lab...throughout the 23-base sequence. Double mismatches ablated Cpf1 activity, except when they were present in...future edits. In contrast, since Cpf1 cleaves relatively far away from the PAM, NHEJ might retain the ...mechanism. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Many research ...

  25. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...genetic tool for researchers to conditionally manipulate gene expression in vivo using site-specific recombination...orientation, the SSR will excise the DNA fragment. By manipulating the number, orientation, and type of target ...morphology, and circuit mapping of a targeted neuronal population.    Figure 2 (modified from Schnutgen...study the physiology or behavior of certain cell populations.  Acknowledgement:Through a partnership with...

  26. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    ...where I gained invaluable experience working on and later chairing the Advocacy Committee. I’m now furthering...more. Having met many people at that conference, I later became involved with the NPA beyond the meeting....more productive and makes it easier to build relationships. Nonetheless, the committees would not function..., I had to learn how to develop new working relationships with these individuals who often had different...

  27. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ... need to deliver the Cas protein altogether. Platforming AAV Interest in interrogating and mapping the...glioblastoma, and generate programmable screening platforms for mouse tissues. Your AAVancement could be next...highly scalable cell type-specificin vivoscreening platform. bioRxiv (Cold Spring Harbor Laboratory). https..., Kalamakis, G., Lischetti, U., Jabaudon, D., & Platt, R. J. (2023). Transcriptional linkage analysis ...

  28. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...2024). Chromatin context-dependent regulation and epigenetic manipulation of prime editing. Cell. https:/...primer binding site and a reverse transcription template (RTT). The design of the pegRNA has been shown...editing for precise and highly versatile genome manipulation. Nature Reviews Genetics, 24(3), Article 3. ...

  29. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...Thursday, or, at the latest on Friday. It’s best to split cells before the plates are more than 80-85%...Benjamin Franklin)! It’s good practice to label your plates and/or flasks with your name, date, the cell line...disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied phycology27.6...

  30. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...heartbeat – a muscle contraction – is directly related to the electrical impulse of the cardiac cells....propagated through multiple cells to do things like translate perceived signals to physical actions. Understanding...serotype specificity during delivery (more on this later). Chemigenetic sensors take a hybrid approach and...JF). The voltage-dependent changes in rhodopsin modulate the degree of quenching of the fluor via FRET....

  31. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At... taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To alleviate...guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you ...the standard information that goes into a 96-well plate and comes out of it. Biotech companies are known...

  32. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    ...of a 5-6 day old human blastocyst [6]. They are isolated mostly from abnormal and/or supernumerary preimplantation...Characteristic ESC ASC Self-renewal High Low once isolated from the human body Usage Universal. Can generate...concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation rejection...when used therapeutically High Low (if ASCs are isolated from the patient) For many years, ESCs or ASCs...

  33. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology...full-color images of proteins. But they are still flat. To truly appreciate the 3D structure of these proteins...   He then said “I hadn’t realized how perfectly flat the complex is on this surface”….where the complex...cleavage”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting...

  34. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ...protein or its role in a disease, researchers often isolate proteins of interest and examine them using biochemical...protein, it's important to validate its function relative to the untagged, wild-type protein. You may, for...the protein of interest and then analyzing the relative colocalization of their fluorescent signals and...One should also be aware of additional problems related to the properties of FPs themselves. These can ...

  35. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...institutions recently reported their inability to recapitulate the results of Gao et al 2016 in a letter to...NgAgo DNA guide is inexpensive and can be 5’-phosphorylated in house using a simple PNK reaction or can... the instability of the supposedly alien 5’-phosphorylated ssDNA guides, which might be rapidly degraded... an ssDNA guide. It is possible that, during translation, only a small fraction of nascent NgAgo protein...

  36. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...double-stranded break. HDR copies the sequence from a repair template with flanking sequence homology for error-free...systems, and NHEJ will be favored even if a repair template is present. HDR also presents a cloning dilemma...CRISPR gene editing. In organisms where HDR is downregulated, MMEJ represents another method for making targeted...’s become clear that the power of this editing platform lies in the diversity of nucleases and their applications...

  37. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ... up after trying out the new protocols.io beta platform for sharing, annotating, and storing life science...molecular cloning protocols, including making LB agar plates, restriction digest of plasmid DNA, and bacterial...bacterial transformation. And more plasmid-related educational resources can be found in our Plasmids 101 blog...the repository. These protocols show up on the related plasmid pages and can help scientists use these...

  38. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started...gaps in the nucleotide sequence, you can run a Translated BLAST (blastx) search to check for expected ORFs..., due to the additional searches involved in translating the nucleotide sequence into all six possible...choose for your sequencing result and can easily translate it, we recommend using blastp over blastx. The...

  39. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...field to a grinding stop and only now, 2 decades later, is it finally back in the limelight. On December...and severe visual impairment that faithfully recapitulated the human condition. Luck brought all these ...X-linked retinitis pigmentosa, achromatopsia, and age-related macular degeneration are ongoing and some will ...capsid variants, designing expression cassettes to regulate and optimize transgene levels, and large scale...

  40. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...would convert adenine to inosine, which the translational machinery treats like guanine. This RNA editor...would permit point mutations in RNA which could recapitulate or rescue known pathogenic alleles, or introduce...ADAR will preferentially edit an adenine if the template has a cytosine mismatch at that position. PspCas13b-ADAR2DD...RNA Editing Capabilities through RESCUE Two years later, the Zhang lab expanded the RNA editing toolkit ...
Showing: 521 - 560 of 974 results