We narrowed to 798 results for: Cre
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TypeBlog PostPublishedJune 21, 2016, 2:30 p.m....including easier automation, higher throughput screening, and a lower requirement for the amount of DNA...transgenic animals may choose Southern blotting to screen founder transgenic animals for the inserted transgene...DNA sequence content by comparing the fragments created by different restriction enzyme digests. Incubating...specific fragments of a DNA sequence of interest to create a physical map of the DNA. Starting in the 1970s...below), restriction mapping may still be used to screen multiple colonies before selecting a smaller subset...allowing for fluorometric-based detection. This increased sequencing speed ultimately leading to what are...
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Providing Plasmids to Researchers in Developing Countries: Addgene and Seeding Labs Team Up
TypeBlog PostPublishedApril 5, 2016, 2:30 p.m....donors to supply surplus lab equipment to carefully-screened research and teaching departments at universities.... Like Seeding Labs, Addgene also feels it’s incredibly important for scientific breakthroughs, reagents...scientific community and are always taking steps to increase plasmid distribution and disseminate knowledge... we need to expand our efforts to find new and creative ways to reach developing countries. As part of... eliminate much of white space on this map and increase plasmid sharing across the globe! We hope that... -
The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing
TypeBlog PostPublishedMarch 10, 2015, 2:25 p.m.... associated hybridoma cell lines. The DSHB was created by the National Institute for Child Health and ... which are each produced by a single B cell. To create a monoclonal antibody cell line, myeloma cells ...individually isolated and expanded. The clonal cells are screened to identify hybridoma lines, each producing a ...and protein tags (c-MYC and HA). We have also created the Monoclonal Antibody Research Institute dedicated...partially humanized rabbit antibodies that exhibit increased avidity to high profile antigens; and 4) generating... -
More Data for You: Find Articles Citing Addgene Plasmids
TypeBlog PostPublishedJune 16, 2015, 2:15 p.m....use the dropdown to increase the length of the list (purple oval in the screenshot below). You can also...plasmid page. Check out the purple arrow in the screenshot below to see what I mean. Additional features...list of papers citing your deposited plasmids Increased visibility of your plasmids in the "Associated...listed in the "Articles Citing This Plasmid" list increasing your work's visibility Help us improve our citations... -
Generating Mouse Models Using CRISPR/Cas9
TypeBlog PostPublishedJuly 12, 2016, 2:30 p.m....specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...knock in models). CRISPR mouse model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide...selection offered by mouse ESCs. For example, we often screen only 15-25 mice when generating knockout models...for generating mouse models Although CRISPR is incredibly useful for generating mutations by NHEJ and generating...CRISPR, one must beware that not all gRNAs are created equal. Some work better than others. There are ...Mice with Extensive Gene Humanization (25-kbp) Created Using Embryonic Stem Cell/Blastocyst and CRISPR... -
Science as Art
TypeBlog PostPublishedMarch 2, 2015, 3:13 p.m....information on black screens, but not to know even the life cycle of the creature they deal with.” So ...Wildlife Magazine. It is easy to see how these incredible images are attracting so much attention. When...efforts, as the subject matter is seen to have increased value and is appreciated by a wider audience for... by holding scientific art competitions where creations can go on display for all to enjoy. The largest... -
CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]
TypeBlog PostPublishedFeb. 18, 2015, 3:09 p.m.... Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell lines. Below... paired DSBs. This method allows for deletion screening by conventional PCR. Biallelically disrupted cells...available strategies by which biologists can imagine, create, and examine novel genetic configurations. Watch...transfected cells via FACS, primer validation, and screening by PCR for biallelic deletion clones: ... and delivery, as well as their techniques for creating deletions, identifying these deletions, and finally... -
What's New in CRISPR - September 2019
TypeBlog PostPublishedSept. 10, 2019, 12:56 p.m.... your research! This time: GeneWeld vectors to create knock-ins CasX Drug inducible CRISPR/Cas9 activation...CRISPR knockout libraries for cancer research Creating knock-ins with GeneWeld vectors GeneWeld from ...Essner’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins of fluorescent proteins, fusion proteins...library with SpdCas9 to perform pooled CRISPRi screens to identify essential genes and phage host factors...metastasis cohorts and a prior mouse metastasis CRISPR screen. The Chen Mouse Tumor Suppressor Gene CRISPR Knockout... -
Quick Guide to All Things Lentivirus
TypeBlog PostPublishedMarch 21, 2017, 2:30 p.m....lentiviral vector system. Viral vectors have been increasingly popular in fundamental and applied research ...replicate. However, Mother Nature found a way to re-create a functional virus. Indeed, by testing several ...the packaging plasmid forming a vector that could create replication competent virus. Even though this system...non-dividing cells and so researchers set out to create a better, safer system. Unlike the MoMLV, lentiviruses...vectors have been established so far, with safety increasing in each generation. The 1st generation of lentiviral...protocols suggest adding sucrose to the supernatant to create a gradient of density that favours virus precipitation... -
Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
TypeBlog PostPublishedMarch 7, 2017, 3:30 p.m.... MOIs. 2. Create a monoclonal Cas9-expression cell line. Especially if you’re doing a screen. I made a...who did not tell you that essential thing ask incredulously, “You seriously didn’t add 3 microliters of ...is bad news for repeatability and bad news for a screen. Because Addgene viruses are provided at a specific... vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014...Additional Resources on the Addgene Blog Genome-wide Screening Using CRISPR/Cas9 Deliver Multiple gRNAs at Once... -
When Fidelity Matters: A frank discussion about ligase fidelity
TypeBlog PostPublishedMarch 6, 2018, 1:49 p.m....reliably detect single-base differences. The original creators of this technique visualized the ligation product...quencher pairs or qPCR-based methods to greatly increase the sensitivity of detection (14-20). LDR has ... 27). This methodology has been used to rapidly screen buffer conditions for Taq DNA Ligase and their ...electrophoresis (CE) (see below). This method allowed screening of all possible base combinations (Watson-Crick...of a 96-well plate, allowing 6 conditions to be screened per plate. The results indicated that the optimal...optimization method has been used internally at NEB to screen additional ligases, conditions, and formulations... -
9 tips for a successful postdoctoral experience
TypeBlog PostPublishedOct. 9, 2018, 12:21 p.m.... strategically to increase your rate of productive results and thereby help decrease the time to your ... to achieve your intended next career stage and create a timeline for achieving these goals. Planning ...ways to consolidate or present your data. You can create more detailed or functional models or more helpful...mentor, manage, communicate, and teach The dirty secret about becoming a group leader is that people are... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum, optimization... -
CRISPR 101: Drug discovery and CRISPR
TypeBlog PostPublishedMay 4, 2023, 1 p.m....CRISPR technology has brought drug and genetic screens together and expedited the drug discovery pipeline...Alternatively, a disease model can be generated, and drugs screened for a change in pathology. If the disease pathology...accelerate the drug pipeline. High-throughput CRISPR screens can be used to inhibit, activate, or knockout many...or an entire pathway of possibilities greatly increases the odds of finding a quality drug downstream....identified several candidate inhibitors of gene X in a screen. You generate a gene X knockout in a cancerous ... -
Plasmids 101: Gene, enhancer, and promoter trapping
TypeBlog PostPublishedMay 2, 2023, 1 p.m....isolated and screened. Depending on the marker, traps can be used as high-throughput screens to introduce...models. Why traps are useful Traditional genetic screens employ various on/off switches to measure the function...circumstances, trap methods may be the preferred screening option. Promoter traps Promoter trap systems ...101: Using Transposons in the Lab Plasmids 101: Screens vs. Selections Plasmids 101: Knockout /Knock-In... -
Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!
TypeBlog PostPublishedJuly 18, 2017, 12:41 p.m....activities (1). She has developed a fast endophyte screening system and plans to employ the light‐inducible...expert. In order to preserve Cassava yields and to increase the potential for this drought tolerant plant,...significant proteins. In this project she aims to create transgenic yeast as an alternative source of essential...plasmids. The recombinant Cry proteins will then be screened for toxicity against local insect pests. By developing... use of chemical pesticides may be avoided or decreased. Biosensors for herbicide detection Dr. Rodrigo... -
Addgene at the ASM Microbe 2016 Conference
TypeBlog PostPublishedJuly 7, 2016, 2:30 p.m....CRISPR screens symposia outlined the fantastic work this Broad Institute lab is doing creating well-optimized...well-optimized CRISPR screening technology. Addgene got several shout-outs for making popular screening libraries ...presence in the microbiology community. For our first concrete steps into this field, we’ve curated microbiology... -
New Viral Vectors - Winter 2025
TypeBlog PostPublishedJan. 28, 2025, noon...pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 AAV2 Recombinase James M. Wilson New serotype pENN.AAV.hSyn.Cre.WPRE.hGH AAV2 Recombinase....GFP-Cre.WPRE.SV40 AAVrg Recombinase James M. Wilson New serotype pENN.AAV.CamKII 0.4.Cre.SV40 AAVrg... viral prep AiP14825: pAAV-AiE0441h_3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN4825) AAV-PHPeB Recombinase... -
Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools
TypeBlog PostPublishedSept. 18, 2018, 1:33 p.m....cobalamin fusion to the riboswitch results in increased fluorescent signal. Prior to the development of...developing a new imaging-based bacterial colony screening platform, Oliver Griesbeck’s lab set out to engineer...the red end of the spectrum (above 600 nm) and increasing the brightness. Arne Fabritius and colleagues...Listen to the base editing podcast segment The increasing popularity of single base-editors has resulted...cytidine deaminases like APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC... -
Hot Plasmids: FRET-Based Biosensors
TypeBlog PostPublishedMay 6, 2014, 1:07 p.m....large-scale functional screen in bacteria. Sensor variants were then refined by a secondary screen in rat hippocampal...protein (CREB) in living cells. “Now we are making this really good,” Griesbeck said. “We know CREB is really...10.1038/nmeth.2773. (PubMed) Friedrich et al. Imaging CREB activation in living cells. J Biol Chem. 2010 Jul... -
Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins
TypeBlog PostPublishedApril 25, 2017, 2:30 p.m....photoconverts predominantly to the anionic form, causing increased fluorescence when excited with 488nm blue light...which allows PA-GFP to exhibit up to a 100-fold increase in fluorescence when illuminated with blue light...performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry (E26V / ...photoswitching. In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't become...the absence of photoactivation resulting in an increased signal to noise ratio. PA-FPs with higher contrast...super resolution microscopy, PA-FPs have helped create several new imaging methods, including photoactivation-localization...