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TypeBlog PostPublishedJan. 21, 2020, 2:30 p.m....CRISPR screen data are also available via comprehensive pre-compiled downloads and via a full-featured REST... to stably archived large-scale datasets by computational biologists. Figure 1: The BioGRID Open... from humans and other species. By accurately compiling genome-wide datasets, which are often obscured...
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Plasmids 101: Positive and Negative Selection for Plasmid Cloning
TypeBlog PostPublishedAug. 22, 2019, 12:43 p.m....synthetize all compounds needed for growth, or auxotrophy, the inability to synthesize a compound needed to...an essential amino acid), and a plasmid that complements the auxotrophy (i.e., synthesizes the critical...yeast prototrophy kit containing 23 plasmids to complement auxotrophy. Negative selection: Cells that have... -
Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!
TypeBlog PostPublishedJuly 18, 2017, 12:41 p.m....projects spanning from the identification of plant compounds and insecticidal proteins to the development of... reside within living plants and that produce compounds with various antimicrobial activities (1). She... newly identified proteins and enzymes. These compounds will later get tested for their potential agricultural...alternative source of essential omega-3 fatty acids complementing vegetable oil, a somewhat poor natural source... -
An Addgene Summer Internship
TypeBlog PostPublishedAug. 8, 2024, 7:22 p.m.... had to get used to the workspaces, as well as complete the lab safety and cybersecurity training before...project is a major project given to each intern to complete and present on. Figure 1: Addgene's...size should be a max of 500 bp (preferably). I compiled a list of three options from the Addgene catalog...Toj-Villagran is a rising senior in high school who just completed their summer internship at Addgene. They plan ... -
An Addgene Co-op: The Intersection of Fun, Friendship, and Work
TypeBlog PostPublishedJune 29, 2023, 7 p.m.... to complete a six month work experience called a co-op to receive a degree. Northeastern compiles a list...Thankfully, my managers set aside time for me to complete an online social media certification on social...petting dogs, and winning the office Just Dance competition. Special thanks to my manager, Sr. Marketing... -
CRISPR 101: Cas9 Nickase Design and Homology Directed Repair
TypeBlog PostPublishedMarch 15, 2018, 12:59 p.m....question is: how should the gRNAs be oriented in comparison to each other? The gRNAs must target different...different strands to create a DSB, but this can be accomplished with either a PAM-in or PAM-out orientation....preferences using HEK293 cells. They started by comparing D10A and H840A total editing efficiency in PAM-in...donor with 40 bp homology arms. The HDR efficiency compared favorably to using WT Cas9 with the left gRNA ... -
Tips for Improving Your Next Manuscript
TypeBlog PostPublishedNov. 22, 2016, 3:30 p.m....t guarantee a publication. There is an art to composing manuscripts and the ASM Scientific Writing and...writer, I was unsure if I should possibly take a composition class in the English Department. He suggested... coming to the writing workshop in D.C. had to compose and submit a manuscript that they were working ...portion of the paper will be generated. After you complete a publication quality figure, produce the figure... -
Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation
TypeBlog PostPublishedApril 23, 2020, 1:15 p.m....platform that allows us to provide scientists with complete plasmid sequences. After launching our viral service...present SMRT sequencing only works for self-complementary AAVs and the TA-based ligation and tagmentation...molecules in the DNA extract associate with complementary strands, enough double-stranded species exist...adeno-associated virus particles containing complementary polynucleotide chains. J Virol 9(2): 394-396... -
Summer SciComm Series: Master of SciComm
TypeBlog PostPublishedJuly 21, 2022, 1:15 p.m....and not solely to ensure a profit. Often, these companies believe they possess something that will truly...the public’s perception of head injury risk. In completing a master’s thesis in science communication, I... I had the chance to carry out a comprehensive review of the literature informing the notion of “heading... Department of Science Communication where he completed a master's degree in 2020, his research explores... -
Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring
TypeBlog PostPublishedNov. 12, 2019, 2 p.m....volunteered to do the sampling and I happily agreed to accompany him to YNP the following year. Geoff Schiebinger...microfluidic parallelization to sequence this highly complex hot spring sample. The sample was separated into...cells each for sequencing, allowing for lower complexity with higher throughput and rigorous assembly....ThermoCas9 from the van der Oost lab. Phylogenetic comparison indicates that like GeoCas9 and ThermoCas9, IgnaviCas9... -
Plasmids 101: TOPO Cloning
TypeBlog PostPublishedOct. 27, 2016, 2:30 p.m....The technique relies on the basic ability of complementary basepairs adenine (A) and thymine (T) to hybridize...deoxyadenosine (A) at the 3' ends of PCR products. The complimentary “T” in the pair comes from a topoisomerase I-linearized...overnight. 4. Transform TOPO Cloning Reaction into Competent Cells: You can use your standard lab protocol ...transforming your reaction into chemically or electro-competent E. coli - excess salt causes arcing during electroporation... -
Extracurricular Activities for a Strong Science Career Path
TypeBlog PostPublishedJan. 21, 2014, 3:08 p.m....great approach to fully understand and acquire competitive skill sets in a myriad of professions is to engage...Mentorship Leadership Science writing competitions (e.g. Access to Understanding) Writing...Communication Networking Publicity Business competitions Leadership Management Marketing ...opportunity to tailor their career paths for competitive positions both within and outside academia. Use... -
Antibody Validation for Flow Cytometry
TypeBlog PostPublishedSept. 10, 2024, 1:15 p.m....intracellular labeling because the intracellular compartment may contain cross-reacting antigens not present...difference in the efficacy of your antibody. When compared, 4% PFA and 0.1% saponin; 4% PFA and 0.1% Triton...intensity in the KO population (pink histogram) compared to the WT cells (green histogram). The unfilled..., like RNAseq or proteomics, it is possible to compare antibody labeling of different cell types within...expression of the target. In this case, you could compare the antibody’s labeling of the cell lines to the... -
Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools
TypeBlog PostPublishedDec. 10, 2019, 1:58 p.m.... the vascular wall. ApoB, the main structural component of atherogenic lipoproteins, shuttles lipoproteins...Thus, the bEFPs bind the UAS via the dCas9-gRNA complex which allows transcription from the promoter. ...should serve as both a powerful tool for generating complex regulatory systems and further the study of natural...hypothesizes that this may be due to different lipid compositions of C. elegans membranes, which might beneficially... -
Creating accessible biology activities in schools with BioBits
TypeBlog PostPublishedAug. 2, 2018, 12:56 p.m....cellular level. The reason for this is cost and complexity – anyone who works in the lab knows how expensive...expensive tiny amounts of reagents are, how complicated (and again, expensive) the different machines are...kits based on freeze-dried, cell-free (FD-CF) components. Making proteins outside of a cell - cell free..., feel free to check out one of our kits and accompanying curriculum to see if any of the activities fit... -
Plasmids 101: Gram Negative and Positive Bacteria
TypeBlog PostPublishedAug. 2, 2022, 1:15 p.m.... bacteria is due to differences in cell wall composition. Cell wall structure Gram positive bacteria, ...is added, it interacts with the CV+, creating a complex within the peptidoglycan layer, staining the cell...the peptidoglycan layer, trapping the large dye complex created by the interaction of CV+ and I- in the...type of bacteria it is being used in, and how components of the plasmid should change based on bacteria... -
Using CRISPR/Cas9 to Edit Disease Out of the Genome
TypeBlog PostPublishedJune 23, 2022, 1:15 p.m.... bulk of which you can find below. CRISPR for complex genetic diseases As CRISPR technology becomes increasingly...using it to investigate potential therapeutics for complex genetic diseases. So we’re revisiting this post...look at CRISPR’s potential for use in treating a complex genetic diseases, Alzhiemer’s disease (AD). Troy...fibrosis (8). The difference with CRISPR/Cas9 in comparison to other genome editing techniques is its ease... -
Visualizing Translation at the Single Molecule Level
TypeBlog PostPublishedAug. 1, 2017, 1:15 p.m....time. Both are helping researchers explore the complexity of translational control in cellular physiology... processing to avoid disrupting ribosome-mRNA complexes. Recent work has adapted this technique to monitor...color switching, TRICK requires the following components: a GFP protein that binds to the coding region... with mRNAs usually translated in a specific compartment. Despite these limitations, these techniques ... -
How Dry Ice Affects Viral Vector Sample pH & How to Avoid It
TypeBlog PostPublishedJune 20, 2017, 2:30 p.m....liquid solution and pH paper to measure the pH. We compared the dry ice-stored samples to controls that were...ice for 72 hours and then vented the tubes by completely removing the caps, incubating the tubes in an...and then recapping the tubes prior to thaw. We compared vented and non-vented tubes to controls that were...various recovery periods between 0 and 96 hours. We compared these samples to controls that were only stored... -
Addgene Depositors Get More Citations
TypeBlog PostPublishedApril 20, 2017, 2:30 p.m....plasmids as if they comprised 10 percent. Based on this new re-weighting, we can compare citations where ...Pamela McA'Nulty and Zachary Tribbett for helping compile the data used in this work. Samantha Zyontz is... topics including biotech and research tools, computing performance, and randomized control trials in ...