Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 731 results for: Sis;

Showing: 441 - 480 of 731 results

  1. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ... well received, understand that people can be resistant to change. Ask your peers or your superiors to...to explain their rationale when they’re resistant to new ideas. If their answer is “we’ve always done ...

  2. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...environmental pollutants, drug discovery, and disease diagnosis. As biosensors, aptamers hold a few important...largely in vitro; Easy and reproducible commercial synthesis (those of us who have used antibodies for diagnostics...

  3. Hot Plasmids - November 2023

    Type
    Blog Post
    ...Susanna Stroik The Liu lab recently used phage-assisted continuous evolution and protein engineering to...  Doman, J.L., Pandey, S., et al. (2023). Phage-assisted evolution and protein engineering yield compact...

  4. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...screening, followed by sequencing and further data analysis. Based on Figure 31.5.2 from https://pubmed.ncbi.nlm.nih.gov...scripts provided by the depositing lab for data analysis.  I'm often asked whether this step can be skipped...

  5. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...however, the protein integration host factor (IHF) assists with the process: it bends the DNA to redistribute...with a beneficial function (usually antibiotic resistance).  In the lab, low copy plasmids may result in...

  6. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...activators The simplest of the dCas9 activators consists of dCas9 fused to a single transcriptional activation...Mediator complex, generally the strongest activator, consists of a dCas9-VP64 fusion and a modified gRNA that...

  7. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...alternative tRNAs Codon optimization With low cost DNA synthesis, one of the primary ways researchers solve the...created codon optimization algorithms and DNA synthesis companies such as IDT and GenScript host online...

  8. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...image shows that the vast majority of the vectors consist of full particles (white arrowhead) relative to...Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense...

  9. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...be made easily and cheaply using oligo library synthesis. Prior to Cas9 activators, similar tools were ...function (Pan et al., 2018). cDNA libraries, which consist of plasmids that over-express coding sequences ...

  10. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target effects of CRISPR/Cas-derived RNA-guided... et al. "Feasibility for a large scale mouse mutagenesis by injecting CRISPR/Cas plasmid into zygotes....

  11. The Challenges of Cell Culture

    Type
    Blog Post
    ...provenance (this is the history of the cell line consisting of records of the origin and propagation of the...Transition Is Required for Acquisition of Anoikis Resistance and Metastatic Potential in Adenoid Cystic Carcinoma...

  13. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...modification of interest, ordering them from a gene synthesis company such as IDT or Genscript, and cloning ...using adeno-associated virus (AAV)." Chromosomal Mutagenesis. Humana Press, New York, NY, 2015. 75-103. PubMed...

  14. Five Popular Model Organisms

    Type
    Blog Post
    ... set of mutations that made cancer cells more resistant to chemotherapy. You can find these plasmids at.... Thus yeast can be used to study the molecular basis of human diseases that are caused by a specific ...

  16. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ..., Y., & Peng, C. (2018). Overview of MicroRNA biogenesis, mechanisms of actions, and circulation. Frontiers...Ørom, U. a. V. (2023). Recent progress in miRNA biogenesis and decay. RNA Biology, 21(1), 36–43. https:/...

  17. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ... transmitted through the integrin, where it is resisted by the ligand embedded in the extracellular environment...development, extended-open integrins enable tissue morphogenesis, organ development, and the shaping of the extracellular...

  18. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... et al. 2022). Lactate is a key metabolite in glycolysis, so this sensor will be a valuable tool to study...lifetime imaging. B) LiLac responses are highly resistant to pH changes. Image adapted from Koveal et al...

  19. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ... steps: 1. Define your timeline Start with your thesis in mind. Imagine the kind of data your dream discovery...consultant and coach in Boston, MA. He specializes in assisting research scientists create productive and manageable...

  20. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...advantage of CRISPR epigenetic tools is their persistence and inheritance. CRISPR activators and repressors... culture. However, DNMT3A-induced methylation persisted throughout a 100 day experimental period, as this...

  22. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...Aschauer, D. F., Kreuz, S., & Rumpel, S. (2013). Analysis of Transduction Efficiency, Tropism and Axonal...Hosokawa, Y., & Konishi, H. (2014). A comparative analysis of constitutive promoters located in adeno-associated...

  23. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...generate more signal from low abundance targets. Analysis/Presentation Once you’ve captured your images,...imaging approach for deep phenotyping and spatial analysis of cells in complex tissues. Proceedings of the...

  24. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...be used for numerous applications, including transgenesis, gene-trap screens, and gene editing. How does... Verma, “Functional Gene Correction for Cystic Fibrosis in Lung Epithelial Cells Generated from Patient...

  27. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    ...protein tags. The basics These tags most often consist of an enzyme that has been mutated to block its...Labeling Technology for Cell Imaging and Protein Analysis. ACS Chemical Biology, 3(6), 373–382. https://...

  30. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...the complexity of the screen. For example, the analysis becomes easier, the number of cells needed is ...add value if you are addressing a well-defined, hypothesis driven research question. Examples of custom ...

  31. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...distinguish different target proteins in your flow analysis, you'll need to use fluorophores that attach to...you can separately detect these markers in your analysis. Lastly, there are fixable dye stains that react...

  32. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...contributed by guest blogger James D. Fessenden, an Assistant Professor at Brigham and Women’s Hospital. Biochemists...Fessenden! James D. Fessenden is currently an Assistant Professor in the Anesthesia Research department...

  34. Pouring LB Agar Plates

    Type
    Protocol
    ... carry one or more antibiotic resistance genes, which confer resistance to a specific antibiotic to the...to be resistant to the antibiotic. On the second plate, streak out a strain that’s not resistant to the... be resistant to the antibiotic, (+) indicates that the tested strain is supposed to be resistant to the...Negative Result 3: Only the Non-resistant Strain Grows If only the non-resistant strain grows, it’s likely ... carrying them. The presence of an antibiotic resistance gene on a plasmid allows researchers to easily...for the selection of only those bacteria with resistance to that antibiotic - usually conferred by a plasmid... plasmid carrying the antibiotic resistance gene. The following protocol will allow you to make your own...

  35. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ... Protocols Agarose Gel Electrophoresis Agarose Gel Electrophoresis You may also like... Restriction...Standard protocol for performing agarose gel electrophoresis, including tips to improve resolution and separation... Agarose Gel DNA Ligation Introduction Gel electrophoresis is the standard lab procedure for separating...pairs) for visualization and purification. Electrophoresis uses an electrical field to move the negatively...protocol video below to learn how to perform gel electrophoresis. Equipment Casting tray Well combs Voltage ...solidified, place the agarose gel into the gel box (electrophoresis unit). Fill gel box with 1xTAE (or TBE) until... short DNA fragments is polyacrylamide gel electrophoresis (PAGE), which is typically used to separate...

  36. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...became automated with the discovery of a heat resistant DNA polymerase from the thermophilic bacterium...several cycles of denaturation, annealing, and synthesis. After the PCR is complete, the product can be...be verified based on size by gel electrophoresis . Protocol Video Watch the protocol video below to learn...temperature step-wise by 1-2°C. The rate of DNA synthesis is ~1-2 kb/min. The extension time can be adjusted...of DNA that we are interested in amplifying for analysis and manipulation. It is essential that the template...cations can also be used for PCR-mediated DNA mutagenesis. A higher cation concentration increases the ...that are starting points for DNA elongation and synthesis. Taq DNA polymerase: Special DNA polymerase that...

  37. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...your restriction digest results. Equipment Electrophoresis chamber Pipetman Reagents Liquid DNA aliquot...manufacturer's instructions) Gel loading dye Electrophoresis buffer Pipet tips Procedure Select restriction...sequence (and where they will cut), use a sequence analysis program such as Addgene's Sequence Analyzer . ...visualize the results of your digest, conduct gel electrophoresis . Tips and FAQ Restriction enzymes MUST be ...Dam or Dcm methylase positive strains will be resistant to cleavage at certain restriction sites. See ...diagnostic digest), you can create a "Master Mix" consisting of all of the reaction components except for ...each tube. This will save you time and ensure consistency across the reactions....

  38. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...separated by denaturing polyacrylamide gel electrophoresis alongside serial dilutions of a standard antibody...Run SDS-PAGE and stain gel Day 1 or later: Image analysis Video Watch this instructional video to learn ...pipette Pipette controller Microcentrifuge Electrophoresis chamber Power supply Rocking platform Fume ...the bottom of the gel. Place the gel in the electrophoresis chamber and secure it. Note: Check the manufacturer...make quantifying the gel easier. Cover the electrophoresis chamber and attach to a power supply. Run the.... Turn off the power supply and unplug the electrophoresis chamber. Remove the gel from the chamber. Use...Addgene QC and would not be used. Section 3: Image Analysis Use ImageJ (Link opens in a new window) or a similar...

  39. Western Blot

    Type
    Protocol
    ...imager -80 °C freezer Reagents 1X PBS Lysis buffer e.g., RIPA lysis buffer Microcentrifuge tubes BCA Assay...in appropriate volume of cold lysis buffer. Pro-Tips The volume of lysis buffer will vary depending on...will generally be between 250–1000 µL . The ideal lysis buffer will vary depending on the cellular location...the bands. Tips and Troubleshooting The optimal lysis buffer will vary depending on the sample type and...protein of interest. You may need to try a variety of lysis buffers to find the best for your target. The optimal...

  40. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis Introduction Restriction enzymes are naturally...and analyze the resulting fragments by gel electrophoresis . The pattern of the fragments on the gel can...enzyme or enzymes that cut your plasmid. Many DNA analysis tools, including Addgene’s Sequence Analyzer ,...to analyze a restriction digest. Equipment Electrophoresis chamber Pipetman Pipet tips Reagents Liquid...manufacturer's instructions) Gel loading dye Electrophoresis buffer Verifying Total Plasmid Size -OR- Insert... tricks see the Restriction Digest and Gel Electrophoresis pages....
Showing: 441 - 480 of 731 results