Skip to main content
Addgene
Showing: 441 - 460 of 486 results
  1. Hot Plasmids - August 2020

    Type
    Blog Post
    ...based antiviral method to degrade RNA from SARS-CoV-2 sequences and live influenza A virus. It was demonstrated...stuck in inclusion bodies. Takayuki Iwaki, Kimiko Hara, and Kazuo Umemura have described a simple method...
  2. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...rate of 10-55% - the PASTE system.     Fig. 2: Schematic of programmable gene insertion with PASTE...and Discovery As part of their quest to find, characterize, and develop new CRISPR systems and technologies...
  3. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...two visible blue or magenta chromoproteins (Figure 2) (Liljeruhm et al., 2018). This system is a nice confirmation...tested their expression under both rhamnose- and arabinose-inducible promoters. Tight control of toxin expression...escape). The rhamnose induction system is well-characterized and quite tight (Giacalone et al., 2006). The...though Khetrapal et al. also demonstrated that the arabinose promoter worked too. Rhamnose induction relies...References Espah Borujeni, Amin, Anirudh S. Channarasappa, and Howard M. Salis. "Translation rate is controlled...
  4. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ... iRFP713, % Oligomeric state Photo-stabilityc, t1/2, s pKa Brightness in HeLa cells vs. iRFP713, % d ...and multicolor NIR imaging.  Small miRFPnano A separate class of NIR FPs, named miRFPnanos, were derived...developed based on mIFP6. Learn more about the characteristics, performance, and applications of near-infrared... found to be a dimer3,15. ND-not determined in parallel assays with other FPs.   We hope that this post...
  5. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...can still record the signal separately for each target.   Figure 2: Examples of simultaneous multiplex...or formalin-fixed, paraffin-embedded tissue sections, methanol- or paraformaldehyde-fixed cell samples...samples, or other preparations. Sequential IF For a sequential IF experiment (Figure 1A), you’ll block, add...different color fluorophore that you can image separately on your microscope. Abcam's sample protocol is...different antibodies, blocking, washing, and sample preparation steps, in order to ensure your antibodies are...Ilangovan, V., Lo, H., Olson, M., Mohamed, S. Y., Sarabipour, S., Varma, A., Walavalkar, K., Wissink, E. M...
  6. Five Popular Model Organisms

    Type
    Blog Post
    ...variety of environmental conditions, and double every 2 hours. Yeast are also the first eukaryotic genome ...as Parkinson’s disease (PD). PD is primarily characterized by protein misfolding that leads to a build-...23594743. PubMed Central PMCID: PMC3703927. Jennings, Barbara H. "Drosophila–a versatile model in biology & medicine...
  7. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ...become a valuable asset yourself.     Fig 2: A diagram of the Thomas-Kilmann Conflict Model, courtesy...favorite books, Michael Ende’s Momo, the title character endears herself to the nearby village (and to ...or how completely different teams are tackling comparable conflicts. Is this level of domain knowledge ...
  8. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    ... Watch Joanne give her "Not" Networking 101 Talk 2. Tailor every talk to the occasion Don’t just plan...come up with clear and concise answers. This preparation will help you avoid giving only yes or no answers...
  9. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...PubMed PMID: 3459152. PubMed Central PMCID: PMC323600. 2. Chester, Nicholas, and Daniel R. Marshak. "Dimethyl...small difference in size, the fragments should be separated on a high percentage agarose gel (~3%). Note that...
  10. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ... from damage (Abe & Lim, 2024).    Figure 2: A) When proteins (blue) interact with the air-water...RvLEAMshort (1:6 molar ratio) with 10 minutes of glutaraldehyde crosslinking: representative micrograph (C)...allowed the authors to determine structures at comparable or higher resolution than using more complex ...buffer additive MgCl2 and the crosslinking agent glutaraldehyde. Adding RvLEAMshort to the sample before plunge...
  11. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...Pluronic-F68 NaCl MgCl 2 KCl Centrifugal filter units (MWCO 100 kDa) Reagent Preparation 1 M NaCl/PBS-MK buffer...Add 5 mL of Buffer B and 2 mL of 5 M NaCl to 43 mL PBS Procedure Preparation and loading of the iodixanol...purification. Workflow Timeline Day 1: Purify Day 2: Buffer exchange and concentration Note: Both steps...buffer Dissolve 5.84 g of NaCl, 26.3 mg of MgCl 2 and 14.91 mg of KCl in 1× PBS in a final volume of 100...at 4 °C. 1X PBS-MK buffer Dissolve 26.3 mg of MgCl 2 , and 14.91 mg of KCl in 1× PBS in a final volume ...need more time, you can alternatively centrifuge for 2 h at 200,000 x g at 18 °C. Carefully take the QuickSeal...interface of the 60% and 40% gradient (see Figure 2) with an 18 ga needle. Place the first microcentrifuge...
  12. AAV ddPCR Titration

    Type
    Protocol
    ... 95 10 2 1 Denaturation 95 0.5 2 50 Annealing/Extension 60 1 2 50 Signal Stabilization 98 10 2 1 Hold ...should decrease by a factor of 2 across the dilutions. In the example below, 2-fold serial dilutions of a ... considered biosafety level 1 but may require BSL-2 handling depending on the insert. Please ensure that...single channel pipette 1–10 µL multichannel pipette 2–50 µL multichannel pipette 20–200 µL multichannel ...20X): 5 µL in 95 µL 1X PCR buffer (1:20) Dilution 2 (20X): 5 µL in 95 µL 1X PCR buffer (1:400) Dilution... DG8 cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction...Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open the QuantaSoft software...
  13. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Activation 95 10 2 1 Denaturation 94 0.5 2 40 Annealing/Extension 60 1 2 40 Enzyme Deactivation 98 10 2 1 Hold ...diploid cells, thus the reason for multiplying by 2. $$V = 2*{copies\ RRE \over copies\ RPP30}$$ Use the viruses...diluted 2-fold serially, the concentration of RRE positive droplets should decrease by a factor of 2 across...Lentivirus is generally considered biosafety level 2+. Please ensure that you are in compliance with your...channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette 20–200 µL multichannel ...Detach cells by incubating with 200 µL TrypLE for 1–2 min. Resuspend cells in 500 µL DMEM complete and transfer... DG8 cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction...
  14. AAV Production in HEK293 Cells

    Type
    Protocol
    ...430825, 175 cm 2 Cellstack 5, Corning 3319, 3180 cm 2 Cellstack 2, Corning 3269, 1272 cm 2 Heat-inactivated...: 50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl 2 Add the following to the 2 L sterile bottle: 1836 mL deionized... solution can be stored at 4 °C for up to 2 months. After 2 months, discard the tube and thaw a new working... mL of PBS. Aspirate PBS and add 2 mL of 0.05% Trypsin/EDTA. Wait ~2 min. Neutralize trypsin by adding...buffer (50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl2) Reagent Preparation DMEM Complete : 10% v/v FBS and 4 mM...Cell-Stack (CS5) (Link opens in a new window) (3,180 cm 2 - the same surface area as 21 x T-175 flasks). Cell...flasks. Workflow Timeline Day 0: Seed cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day...
  15. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...biology 14.6 (2004): 757-764. PubMed PMID: 15582400. 2. Tomkinson, Alan E., et al. "DNA ligases: structure...the ligation junction were represented. Sixteen separate pools were prepared, each with a different complement...applications such as routine cloning or DNA library preparation. However, when a method relies on accurate ligation... 19329793. PubMed Central PMCID: PMC2719376. 5. Barany, Francis. "Genetic disease detection and DNA amplification...PMID: 1986365. PubMed Central PMCID: PMC50775. 6. Barany, Francis, and David H. Gelfand. "Cloning, overexpression...PMCID: PMC208194. 8. Luo, Jianying, and Francis Barany. "Identification of essential residues in Thermus...76.2 (1989): 245-254. PubMed PMID: 2753355. 12. Barany, Francis. "The ligase chain reaction in a PCR world...
  16. Finding Your Perfect Job After University

    Type
    Blog Post
    ...experience in cancer research After graduating with a 2:1 BSc in Molecular Biology (roughly a B average in...really passionate about during my degree. I loved parasitology. However, when I enquired about roles in this...end of my three months, I fully understood why parasitology roles in the field require experience. Working...beginning of 2010, I explored options for going into parasitology but another major factor held me back; I do ...
  17. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...Nutr, 129 (2009a): 477–484. PubMed PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic...Forster, A. C., Skingle, D. C., & Symons, R. H. “Preparation and uses of photobiotin.” Methods in Enzymology...
  18. Fluorescence Titering Assay

    Type
    Protocol
    ...method 1) or the volume of virus (method 2): Method 1 Method 2 $$T = {N*F*D\over V_T}$$ Where: T = Titer...Lentivirus is generally considered biosafety level 2+. Please ensure that you are in compliance with your...Day 0: Seed 293T cells Day 1: Transduce cells Day 2 (am): Remove media, replace with fresh media Day 4...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel... 200–1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container...complete. Mix well by pipetting or inverting. Aliquot 2 mL of cell suspension into each well of the 6-well...conical tubes, VWR 21008-216 Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM...
Showing: 441 - 460 of 486 results