...single-strand DNA incorporation (ssDI). "Easi-CRISPR for creating knock-in and conditional knockout mouse models...Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in mouse models with CRISPR has been challenging...efficiency. Easi-CRISPR uses a long ssDNA template to increase efficiency of insertion and gene replacement. ...

...we found that adding a TEV linker dramatically increased accessibility of the AviTag and HisTag, which ...fusion protein in bacteria. Our initial attempt to create a plasmid based on the biotin ligase BirA backbone... inserting BirA into a pMal plasmid (NEB) thus creating the pMal-T-Avi-His/BirA plasmid now available ...

...cassette allowing a researcher to perform blue-white screening after transformation - self joining of the vector...break down the steps needed for TOPO cloning: 1. Create Your PCR Product: Design standard primers (no need...unpaired 3’ ends in your PCR product. If you need to decrease error rate, please use one of these methods to...

...the community, increasing the impact of the work, and as a collateral benefit, increasing the impact of...Tompson Institute.  References Baldwin, Melinda. "Credibility, peer review, and Nature, 1945–1990." Notes and...

...individual cells. Each cell is labeled with 3-4 tags to increase the diversity of the CellTag library. The fixed...detected over a 10-week period. Their expression decreased over time but they were not silenced.    Find ...retroviral overexpression cocktail yielded a three-fold increase of fully reprogrammed cells based on single-cell...

...University of Alberta and University of Oregon created mTFP1 by transforming E. coli with a gene library...tetramer-breaking mutations. The library was then screened through successive rounds of mutagenesis for the...filter. This combination results in a 2.6-fold increase in signal relative to mCerulean imaged with a ...

...primary antibody and endogenous IgGs in the tissue. Created with biorender.com. Strategies for avoiding...Some labs have found that you can significantly decrease background staining by mixing your primary and...you add your primary and secondary antibodies. Created with biorender.com. Proper tissue processing...

...collection of antibodies available at launch were created by James Trimmer, PhD, of the University of California...commitment to supporting your research, Addgene is creating an array of educational materials to accompany...more useful to everyone over time. Through our incredible community, including your support, we have enabled...

...to select against CRISPR positive plants, but screening these can be challenging and requires specialized...Willoughby   Using RUBY as a selection marker The creators of RUBY highlight its use in crop plants under...as positive selection of transformed plants to decrease the routine usage of antibiotics which may contribute...

...would rather fund excellent people with mediocre ideas than mediocre people with excellent ideas. I am lucky... your potential as an entrepreneur, not your credentials. If you have an idea, don’t wait.  Go for it....

...microvesicles (approx. 40 - 150 nm diameter) that are secreted from the cellular membrane into the culture medium...AAVs (Schiller et al., 2018). Exosomal vesicles secreted from producer cells also contain cellular macromolecules...open new ways of improving gene delivery. They increase the efficiency of transduction, broaden the use...

...to keep the audience engaged through a computer screen. For the Reproducibility for Everyone Workshop,...author of the poster,” he says.Viewing a poster on a screen is challenging, requiring you to zoom in to different...canceled may be disappointing, but there are many creative ways to adapt and take advantage of the new conference...

... the stem cell community – a community largely created due to Yamanaka’s seminal work. It didn't take... during his keynote address, appeared almost incredulous that the field has exploded so quickly, citing...explained that it took 50 people nearly 2 years to create the first stem cell line that could be used in ...

..., described below, are typically employed when creating stable cell lines and are not used for general...Stable transfection A stable transfection is used to create a population of cells that have fully and successfully...culture media until 24-48 h post transfection when creating stable cell lines. Looking for Mammalian Expression...

... vector and insert are (separately) digested to create 10–12 base “chewed back” overhangs, they can be...bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   SLIC also opened up a...transformed into bacteria, which repair the plasmid. Created with BioRender.com. How does SLIC compare to...

...and their GPCR binding partner, Bryan Roth’s lab created an open source resource called PRESTO-TANGO (parallel...parallel receptorome expression and screening via transcriptional output - TANGO) (Kroeze et al., 2015...are ideal candidates for use in chemogenetics. To create chemogenetics receptors, GPCRs were first mutated...

...sequence (pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP ...virus, making it ideal for CRISPR pooled library screening experiments. Find lentiCRISPR v2.(If you’ve peeked...site, a pair of annealed oligos can be cloned to create a chimeric guide RNA. Find pX330-U6-Chimeric_BB-CBh-hSpCas9...

...under cold temperatures, and frigid conditions decrease the potential for microbial contamination. Additionally...Acclimatisation To resolve this impasse, our team decided to create a hybrid system where we could incorporate cold-tolerance...The event also featured a seminar on “Tools to increase scientific reproducibility” by Benoit Giquel, ...

...well-characterized. During this time, we’ve also created a variety of educational resources in an effort...understanding of Addgene’s customer’s needs. Categories We created a categorization of our information in order to...for promoter and serotype, among others. We also created a normalization mechanism to “collapse” variations...

...production. Finally, we are blessed to have an incredibly supportive Board of Directors at Addgene. Our...leadership of Addgene’s co-founder, Melina Fan, we created a new team to push this project forward. The viral...you to the Addgenies who went the extra mile to create this new service. We knew the scientists were waiting...