We narrowed to 790 results for: Cre
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TypeBlog PostPublishedJan. 3, 2019, 1:35 p.m....cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates a nick in the DNA by cutting only a single strand...inhibitor (UGI) minimizes the second outcome and increases the generation of the desired T-A base pair. ...act on DNA for adenine base editing. They first created a defective chloramphenicol resistance gene (CamR...antibiotic resistance. To find such a protein, they created a mutant library of E. coli tRNA adenosine deaminases...into E. coli harboring the defective CamR gene. Screening for surviving colonies and subsequent rounds of...genotyping. If successful base editing causes the creation or destruction of any restriction enzyme (RE) ...
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Using Video to Share Your Science: We Share Science
TypeBlog PostPublishedNov. 3, 2016, 2:30 p.m....humanities, and 32% in the social sciences go uncited. Creative titles and controversial topics can garner some...Article, since the 1980s researchers have been increasingly citing research from outside of their disciplines...interdisciplinary research while at the same time increasing interdisciplinary citations in their own work...instant sharing across multiple other sites), I created a new platform for sharing video abstracts about...effectively.” How to share your science through video Creating an effective video about your research is not ...offers a 5-step process and several examples for creating effective research videos. Karen McKee, also... -
2021 Deck the Lab Winners!
TypeBlog PostPublishedJan. 25, 2022, 2:15 p.m....Saves Christmas! created by the Moore lab at the Van Andel Institute. Krampus was created by Nate Levine...Warrior The Moore lab, who was full of holiday creativity, also receives an honorable mention for their...2021 2nd Place: The Winter and the Falcon Tube created by the team efforts of the Yit Heng Chooi Lab at... December 4, 2021 3rd Place: O! Beaker Tree Created by Joanne Jary from Lab Support at the John Innes... -
CRISPR 101: Homology Directed Repair
TypeBlog PostPublishedJan. 26, 2023, 2:15 p.m....DNA end of the break is resected by nucleases to create a ssDNA 3’ overhang. This will serve as both a ...considerations for designing a repair template to create mutations HDR donors can contain insertable sequences...circular plasmid templates is generally low; to increase the frequency of edits, researchers have designed...complementary ssDNA. Easi-CRISPR was shown to increasing editing efficiency from 1-10% with dsDNA to 25...Frequency of Homology-directed Repair (HDR) for Increased Efficiency of CRISPR/Cas-mediated Transgenesis... -
Evolution of Lab Techniques
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... -
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... -
Viral Vectors 101: AAV Serotypes and Tissue Tropism
TypeBlog PostPublishedJan. 16, 2025, 3:42 p.m..... Figure 1: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive...AAV2, and so on. In addition, researchers have created many engineered serotypes by mixing, matching, ...been shown to transduce skeletal muscle, cardiac, pancreas, smooth muscle, CNS, kidney, and retinal cells...been shown to transduce liver, skeletal muscle, pancreatic, and retinal cells (Issa et al., 2023). AAV9 ...pUCmini-iCAP-AAV9.452sub.LUNG1 Goertsen et al., 2022 Pancreas AAV6, AAV8 Wang et al., 2006 Retina 7M8 Dalkara... J., & Xiao, X. (2006). Widespread and Stable Pancreatic Gene Transfer by Adeno-Associated Virus Vectors... -
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... -
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... -
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... -
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... -
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... -
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 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... -
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... -
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: 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... -
Visualizing Genomic Loci with CRISPR-Sirius
TypeBlog PostPublishedApril 18, 2019, 12:53 p.m....addition to increased brightness, they also found that the signal to noise ratio can be increased from 2-fold...previous CRISPR-based loci visualization tools by increasing gRNA stability. First, they began with the Broccoli...and went through subsequent design iterations to increase stability and minimize misfolding and recombination...label two loci simultaneously, Ma and Pederson created another sgRNA containing the PP7 hairpin sequence... -
The Importance of High Titer for AAV Transductions
TypeBlog PostPublishedMarch 1, 2018, 2:11 p.m....roughly 200 cells. Increase the titer to 1013 resulted in a roughly 6-fold increase in expression [3, ...Figure 2]. Note how expression increases exponentially with an increase in titer. That means that once...to get more AAV in contact with the cell is to increase the AAV concentration. In this flowchart, you ... -
Plasmids 101: Gram Negative and Positive Bacteria
TypeBlog PostPublishedAug. 2, 2022, 1:15 p.m....purple) and Gram negative (red/pink) staining. Image credit: Mark Perkins https://www.flickr.com/photos/occbio...charged iodine is added, it interacts with the CV+, creating a complex within the peptidoglycan layer, staining...peptidoglycan layer, trapping the large dye complex created by the interaction of CV+ and I- in the thick multilayered...positive and negative classification system is incredibly helpful in visualizing differentiations between...What is a plasmid? Plasmids 101: Blue-white screeningPlasmids 101: Common lab E. Coli strains ... -
K. phaffii: Rising to the Occasion
TypeBlog PostPublishedApril 4, 2023, 1 p.m....protein secretion, making harvesting recombinant proteins a cinch. This is because it doesn’t secrete its ...of producing high protein animal feed. Due to increasing prices of methanol and advancements in other ...proteins, making the recombinant protein its major secretion. Beyond sheer protein production, K. phaffii is...Organisms Plasmids 101: Yeast Vectors Tips for Screening with the Yeast Two Hybrid System Additional... -
Plasmids 101: NGS Quality Control for Pooled Libraries
TypeBlog PostPublishedOct. 26, 2017, 1:59 p.m....primers to create 200-300 bp sequencing products using PCR. A diagram of the product we created for the ...plasmid. However, lots of library DNA is needed for screening experiments, so the scale of the DNA prep is larger...rank the plasmids by abundance, scaled to 1, to create a “cumulative wealth distribution” curve, or Lorenz...wishes you luck in your future library use and screening, and if you have any tips, input, or stories of... -
Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond
TypeBlog PostPublishedJune 1, 2021, 1:15 p.m.... lack of credit for small contributions. We strive to eliminate these two barriers, creating a community...with others than by oneself that prompted us to create Crowdfight. Figure 1: Crowdfight is a ...collaboration between researcher and expert; 4) Credit all contributions. Our volunteers are extremely...collaborations can be easily started and easily credited. And nothing stops these one-time favors from ... -
Fluorescent Proteins 101: Luciferase
TypeBlog PostPublishedJune 24, 2014, 3:59 p.m....resulting in a decrease in luciferase activity. This experimental protocol can readily screen miRNAs for ...Peak emission (nm) Size (kDa) Substrate Cofactors Secreted Luminous shrimp(Oplophorus gracilirostris) 455...cell membrane and deliver the substrate; however, secreted luciferase or the use of alternative substrate...pathways can be investigated in a high-throughput screen using luciferase activity as a measure of signaling... -
Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors
TypeBlog PostPublishedFeb. 19, 2019, 1:36 p.m....the potential for increased speed and efficiency through which knowledge is created and shared. Can you...platform. Expanding further beyond the US and increasing Addgene’s international presence would potentially...information, making it available to everyone, that creates more breakthrough and innovation. With any kind...science regulatory law. My role at Suffolk is to help create new programmatic content as well as help our students... -
7 Tips to Secure a STEAM Internship This Summer
TypeBlog PostPublishedFeb. 2, 2021, 2:15 p.m....that approach seems to create competition for your own spot. True! But you create a network of friends,... and simplify access to academic resources, we created STEAMid, for a nationwide database of STEAM internships...with your question(s). This precautionary step increases the likelihood of acceptance to the internship... following graduation. While these suggestions increase your competitive edge, they alone, are no guarantee... -
Kiran Musunuru on the Newest TALEN Genome-Editing System
TypeBlog PostPublishedDec. 10, 2013, 3:13 p.m....sequence desired. This technology allows you to create a pair of designer proteins that will introduce... transfection or electroporation. All you do to create a protein that binds those 15 base pairs is take...genome editing tools allow you to do is greatly increase the efficiency of the genome editing process. ...for investigators without a lot of expertise to create zinc finger nucleases that will bind desired sequences...in terms of the target site. Also, any time you create double-strand breaks, there is a possibility of... -
RaPID Detection of RNA-protein Interactions
TypeBlog PostPublishedAug. 16, 2018, 12:40 p.m....address this need, the Khavari lab at Stanford created the RNA-protein interaction detection (RaPID) method...RaPID. Ramanathan et al. also tweaked BirA* to increase its rate of biotinylation. The improved enzyme...in neural progenitor cells. Knock-down of QKI decreased Zika viral RNA levels by 90%, suggesting that ...Hyperferritinemia-Cataract Syndrome (HHCS), a disease characterized by increased iron serum levels and early-onset cataracts, is...sequences and that loss of binding correlated with increased serum iron levels seen in HHCS patients. These... -
RANbodies: Reporter Nanobody Fusions
TypeBlog PostPublishedApril 10, 2018, 12:56 p.m....antibody and a ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to...is a ~150 amino acid, single domain antibody. To create their RANbodies, Yamagata et al used previously... RANbodies are expressed in 293T cells and are secreted from cells due to their IgGκ signal peptide sequence...them. HRP RANbody staining results in a ≥10-fold increase in signal over staining with a primary/ labeled... -
Human Germline Editing Using CRISPR
TypeBlog PostPublishedAug. 10, 2017, 2:19 p.m....also underwent editing through the NHEJ pathway. Created with BioRender.com. In later characterization...current tools, thus quieting some fears about the creation of “designer babies.” The differences in editing...disorders, pre-implantation genetic diagnosis (PGD) screens embryos to find those that do not carry a given.... claim that CRISPR would still be helpful in increasing viable embryos, especially in the case of older...still be worthwhile for a given disease if it increased the percentage of disease allele-free embryos ... -
CRISPRainbow and Genome Visualization
TypeBlog PostPublishedFeb. 28, 2017, 3:30 p.m....scaffold to enhance complex assembly, thereby decreasing the amount of background fluorescence from unbound...CRISPR to label multiple loci in distinct colors. To create a colorful Cas9 toolbox, Ma et al. turned to SpCas9...was fused to a different fluorescent protein to create three distinct colors. The specificity of these...distinct hairpin sequences produces white light. Created with BioRender.com. With this rainbow of colors...combination, for example, with a far-red FP, would increase the available colors to 15! They also envision... -
The time and cost required to make a plasmid
TypeBlog PostPublishedMarch 23, 2023, 1:15 p.m....Institute has created hundreds of thousands of plasmids, using both mass production genome wide screens and the...vector. The time it takes to make a plasmid could increase dramatically if you are piecing together several...at Addgene, the consumables cost per stab has increased 50% since 2019, while overall inflation of USD...equal to the $155 spent on reagents. Five ways to decrease plasmid production time and cost If you’re short... -
Celebrating 15 Years of Scientific Sharing
TypeBlog PostPublishedJan. 14, 2019, 2:08 p.m....visibility is increasing internationally, thanks to evolving outreach tactics, increasing business development...they see the value in depositing and they are increasingly seeking us out on their own to share reagents... We were and remain ahead of most companies in creating a welcoming, inclusive workplace for everyone....about their dedication, excellent intentions, or incredible drive to succeed. Thanks to all the Addgenies... -
Gendered Innovations: Why Does Sex of the Cell Matter?
TypeBlog PostPublishedJuly 21, 2016, 2:30 p.m....Foundation, and Stanford University—that harnesses the creative power of sex and gender analysis for innovation... in research Governments, universities, and, increasingly, corporations in the US and Western Europe have...decades: “Fix the Numbers of Women” focuses on increasing the numbers of women participating in science...gender analysis and 2) provides case studies with concrete examples of how gender analysis leads to discovery... Environment—a project aimed at harnessing the creative power of sex and gender analysis for innovation... -
The Future of Research Symposium Boston 2015
TypeBlog PostPublishedOct. 20, 2015, 2:30 p.m....these demands? Are there other solutions, such as increasing numbers in Master’s level programs? What would...Maryland College Park, is a strong supporter of increasing the number of postdoctoral scientists at University...School. He has continually promoted the cause of increasing data collection about the scientific workforce... Boston 2015 where we hope to begin developing concrete solutions to the problems looming over the academic...School. In the lab, he uses synthetic biology to create tools to better understand human disease. Outside... -
Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux
TypeBlog PostPublishedSept. 13, 2016, 2:30 p.m....particularly given that this has precipitated increases to the NIH recommended pay scale – used by many...while this is a move in the right direction, the increase notably still does not meet the suggested postdoc...this area, keeping the pressure on for further increases will be challenging, but will be aided by the ... new strategies designed to cope with current increases. These two rulings along with progress over the...School. In the lab, he uses synthetic biology to create tools to better understand human disease. Outside... -
Illuminating Epigenetics with A FRET Based Biosensor
TypeBlog PostPublishedNov. 19, 2015, 3:30 p.m....grandsons that decreased their likelihood of death from cardiovascular disease but increased their risk of... DNA methylation patterns, either large increases or decreases. Hypermethylation of CpG islands in the... units of the FRET pair together and causes an increase in the FRET signal. Subsequent demethylation separates... -
Cloning Mammalian Cells with the Agarose Method
TypeBlog PostPublishedSept. 7, 2017, 12:17 p.m....amount of agarose/DMEM should be proportionately increased). For some cells, add a drop of serum to aid in...up, most will eventually grow within a week. We screen the wells in groups as they become confluent, taking...important to have a functional assay in place to screen your clones. Depending on the protein, we have ...SH-SY5Y cells, as these cells did not seem to form discrete colonies but constantly migrated away from each...either initially, or using secondary biological screens. For example, for cancer researchers, if growth... -
Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles
TypeBlog PostPublishedJune 22, 2017, 2:30 p.m.... to its final destination. As an example, most secreted proteins are produced in the endoplasmid reticulum...golgi and then transported in vesicles via the secretory pathway to their subcellular or extracellular ... they are finally degraded (2, 3). Once you've created your fusion protein, it's important to validate... to make any fluorescent protein constructs you create in your own work available to the Addgene community...Edition. 3. CureFFI.org. Cell Biology 04: The Secretory Pathway. 4. Dunn, Kenneth W., Malgorzata M. Kamocka... -
Antibodies 101: Flow Compensation
TypeBlog PostPublishedMay 16, 2024, 1:15 p.m....needs to be compensated out of APC-Cy7. After increasing the compensation values for APC against APC-Cy7...to be shifted towards the x-axis, so you would increase the compensation value for APC. If the population... to be shifted away from the x-axis, you would decrease the compensation value. Figure 1: Compensation...population needs to be shifted towards an axis, increase the compensation value, whereas if it the population...population needs to be shifted away from an axis, decrease the compensation value. If neither of these helps... -
Antibodies 101: Conventional vs Spectral Flow Cytometry
TypeBlog PostPublishedAug. 3, 2023, 1:15 p.m....detectors for all fluorophores. This allows for the creation of much larger panels, as fluorophores that cannot...they don’t “wash out” the rest of the panel and create spreading error. Conversely, markers with low expression...should not be paired with these dyes, as this would create difficulty when trying to resolve those populations...signature upon antibody binding, leading to discrepancies during unmixing. Additionally, it is critical...spectral flow cytometry has the ability to generate incredible amounts of data from a single experiment. Although... -
Hot Plasmids and Viral Preps - May 2021
TypeBlog PostPublishedMay 18, 2021, 1:15 p.m.... aaRSs in these processes. To create the antibody toolbox, they screened antibody candidates using a synthetic...fourteen aminoacyl tRNA synthetases (aaRS) have been created, characterized, and validated by the Gräslund lab...cross-reactivity, antibody candidates underwent affinity screening to select for slow off-rate binders. The best ...pathways By Gabrielle Clouse Claudia Vickers’ lab has created a novel nanocompartment for transporting proteins...