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TypeBlog Post...fibrillary acidic protein (GFAP) for astrocytes it is important to test it in the brain region of interest, especially...of Transduction Efficiency, Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse... the Addgene blog Plasmids 101: FLEx Vectors Important Considerations When Using AAVs Three Key Considerations...
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Mycoplasma Contamination: Where Does It Come From and How to Prevent It
TypeBlog Post...mycoplasma contamination The best and the most important ways to avoid mycoplasma are by providing proper...viability. Conclusion To sum up, it is always important to prevent contamination rather than eradicating...others when you can and help them understand the importance of preventing contamination. Be aware and cautious... -
Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
TypeBlog Post...of the gene with multiple gRNAs, but the most important is that you could avoid potential off target artifacts...you could run another gel that highlights the important results (for example, if you needed to make a ... Despite the shifting landscape of research opportunities, it’s an exciting time to be in the lab. Based... -
Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2: Reprogramming and Transdifferentiation
TypeBlog Post... adenovirus [5], Sendai virus [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]...insulin-producing β-like-cells [37, 38]. In 1987, Davis et al. reported one of the earliest examples of transdifferentiation...differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl... -
Prime Editing: Adding Precision and Flexibility to CRISPR Editing
TypeBlog Post... DNA to favor the edit Once the prime editor incorporates the edit into one strand, there’s a mismatch...MutSα–MutLα mismatch repair (MMR) complex, to temporarily inhibit this process and promote copying of the...it from hybridizing with the unedited strand. Temporarily inhibiting this complex may allow time for 5′... -
CRISPR Library Amplification
TypeProtocol... mix. Electroporate cells (one at a time for a total of eight electroporations): Electroporator Conditions...and have 1 mL SOC per electroporation readily available for post-electroporation recovery of cells. Ensure...if the recombinant band makes up a significant proportion of the DNA pool. Last Update: August 17, 2023...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents.../or increasing the skewness of the pool. 8 electroporation cuvettes (BioRad, Micropulser, 0.1 cm ) 20 ...convenience but any brand or house- made LB that supports normal growth is expected to work. 50 mL Falcon...DNA-Endura into pre-chilled cuvette. Pulse. Caution Electroporation involves the use of high voltages, please use... -
CRISPR 101: Cytosine and Adenine Base Editors
TypeBlog Post.... They also mutagenized the cytidine deaminase portion of the base editor to create SpCas9 base editors...off-target activity (Richter et al., 2020). This is important because ABEs have generally been slow, meaning...editing for plants Read about four base editor reporters Additional Resources on Addgene.org Catch up... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post...the phage recombineering machinery are first electroporated with pCas9. Then, pCRISPR is introduced along... a mammalian cell line (Qi et al., 2013). Two important features of this early system are its regulatable...Euler CW, Jiang W, Nussenzweig PM, Goldberg GW, Duportet X, Fischetti VA, Marraffini LA (2014) Exploiting... -
Trends in CRISPR and SynBio Technologies
TypeBlog Post...reflections on the program are here. This was a great opportunity for Addgene to present our own data on plasmid...in advancing this field and how we are able to support rapid progress in all areas of research. We welcome... -
New FLIS Plasmid (April Fools 2022)
TypeBlog Post...more commonly known as fruit flies, have been an important animal model for research, particularly genetics...The Frequency Light Interval System, or FLIS, incorporates a bioluminescence protein into the chitin that... -
Twenty Years of Sharing Science!
TypeBlog Post...India, and Brazil. We've also created a robust support system for scientists using materials in our repository...repository, from our customer and science support to our efficient and easy tech transfer process to our... -
Plasmid Cloning by PCR (with Protocols)
TypeProtocol...Product Run PCR to amplify your insert DNA. It is important to use a high fidelity taq polymerase to minimize.... The fidelity of the polymerase becomes more important the longer the expected PCR product is. You should...temperature based on the melting temperature (Tm) of the portion of the primer that hybridizes to the sequence to...some DNA during the gel purification step, it is important to digest plenty of starting material. We recommend...be cut with both enzymes, and therefore it is important that the digest goes at least 4 hours and as long...running a gel for purification purposes it is important to have nice crisp bands and to have space to ...expected. When cloning by PCR, it is especially important to run the product on a gel. This allows you to... -
Affinity Purification of Recombinant Antibodies with Protein A or Protein G
TypeProtocol...446 Benzamide, Millipore Sigma 12072 Antipain, Millipore Sigma 10791 Leupeptin, Millipore Sigma L2884 Aprotinin...monobasic monohydrate, Millipore Sigma 71507-250G Sodium phosphate dibasic, Millipore Sigma S7907-500G 1 ...that sharing the full details of our protocols supports reproducibility and accelerates science. Here,...information is solely for transparency intended to support reproducibility in science. Safety Warnings Sodium...1 M Trizma hydrochloride solution pH 9.0, Millipore Sigma T2819-1L 250 mL sterile bottle, VWR 430281 0.45... Aprotinin, Millipore Sigma A6279 Zeba Spin Desalting Columns, 7 kDa MWCO, 10 mL, Thermo Fisher 89893 ...89893 Amicon Ultra-15 30 kDa MWCO column, Millipore Sigma UFC903024 50 mL LoBind tubes, VWR 76289-498 Before... -
Imaging Tools and Following a Passion for Basic Biology: Interview with Joachim Goεdhart
TypeBlog Post...biology and how it speeds science. Chapter 8 - The importance of communicating research findings through social... singapore... -
Call for Guest Bloggers
TypeBlog Post...Whenever possible, we love to give scientists the opportunity to share their knowledge. One of our goals is...social media outlets meaning you’ll have the opportunity to interact with and discuss your work with many... -
New Viral Vectors - Spring 2025
TypeBlog Post...to meet the diverse needs of our community and support a wide range of experimental designs. Here are ...help, with over 100 user-submitted viral vector reports. And don't forget to use the link below to keep... -
New Viral Vectors - Summer 2024
TypeBlog Post...synapses distant to the site of injection with high temporal precision. Table 1: New ready-to-use viral ...check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the link... -
Kwabena Duedu on Public Health, Biofuels, and Doing Science in Ghana
TypeBlog Post... well as plasmids from Addgene to bring new opportunities to Ghanaian scientists Click here to listen... Kwabena's goal to build biological science opportunities in Ghana. 46:35 - end : Kwabena's time in Boston... -
The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing
TypeBlog Post...repositories, such as Addgene and the DSHB, play an important role in scientific exchange. In addition to ensuring...provide standardized quality-control and technical support to the community. Thanks to Our Guest Blogger! ... -
Plasmids 101: How to Name Your Plasmid in 3 Easy Steps
TypeBlog Post...identify what your plasmid contains and other important information. Tip: A lowercase "p" is often used...simple piece of information can often convey many important details. Once you know the backbone a plasmid ...