...our guest blogger, Jon Chow! Jon Chow is finishing his immunology PhD at Harvard University. He has had ...three larval stages where larvae molt in order to continue growing in the next larval stage. After the ...the third larval stage, Drosophila will pupate, which takes about one week from the initial deposition of...through their translucent bodies. This is the ideal stage to select for females because you can be certain...

...developed and demonstrated by Manuel Gonçalves and his colleagues at Leiden University Medical Center are...-transformed cells yielded rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding...gRNAS1, and pAdSh.U6.gRNAGFP. Gonçalves says that advantages of AdVs include their episomal nature and very...used, the CRISPR approach based on HDR has the advantage of being a potentially universal treatment (Stephens...CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells. Sci Rep 4:...

... who later shared a Nobel Prize in Chemistry for his work. Since then PCR has become a standard and essential...generating mutated sequences for site-directed mutagenesis.  Quantitative polymerase chain reaction, qPCR...interest  Molecular Cloning Site-directed mutagenesis Molecular Diagnostics qPCR Quantify starting... PCR  Plasmids 101: Colony PCR Site Directed Mutagenesis by PCR Resources on Addgene.org Protocol: Polymerase...

...antibodies against the same target; tagged protein strategies that express a tagged version of the antigen and ...compare staining of the antibody to that of the anti-tag antibody; and immunoprecipitation-mass spectrometry...be variations within an application. Take immunohistochemistry (IHC), for instance. While an antibody may...

... mammalian) Tags (e.g. 6xHis, Flag, StrepII, HA, Myc, SUMO, GST, MBP, CBP, S-tag, TAP tag, TRX, or V5)...pseudovirus (Ctrunc) ID Plasmid Description Mutations Tags Industry Additional Resources Read Addgene's Blog...

...to immunoprecipitate FLAG-tagged Cas9. In the CAPTURE system, Cas9 is tagged with a biotin acceptor site... Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs...system from the Bassik lab, uses an MS2 hairpin-tagged gRNA to recruit a cytidine deaminase and induce... Available modifications include histone acetylation by p300, histone demethylation by LSD1, cytosine ...

...al., 2023). Our thanks to Rodolphe Barrangou for his authorial contributions to, and support for, this... shipped out each year.     Figure 3: Percentage of requests per year for CRISPR plasmids categorized...

...generate one molecule of GFP-tagged LC3 for every one molecule of RFP-tagged LC3∆G. Like the unmodified ...of autophagic substrates inside the lysosome. Historically, methods for studying autophagy focused on counting...

...frequent bouts with Impostor Syndrome, Joe has found his people through meetings and social media. Joe is ...conference hashtag I had, however, started a Twitter account, and was following the meeting hashtag. I noticed...Favorable conditions - Expansion of networks The last stage is Exponential Growth, named after the growth phase..., and the Society for Advancement of Chicanos/Hispanics and Native Americans in Science’s (SACNAS) annual... career scientists, students, and those from historically underrepresented groups. Through such mentoring...

... vivo. Some researchers have used fluorescently tagged DNA-binding proteins to label certain loci, but...Kits Find Empty Backbones with Fluorescent Protein Tags ...the 4D nucleome. Visualizing specific loci has historically been difficult, as techniques like fluorescent...this rainbow of colors, Ma et al. performed sophisticated tracking of multiple chromosomal loci in live...

...lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters, specific time points...plasmids here! Lin D., Li X., et al. (2023). Time-tagged ticker tapes for intracellular recordings. Nature...externally-timed bands of color with HaloTag-iPAK4 by applying different HaloTag dyes and showed they could record...proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching an old antibody....1101/2023.02.07.527548.    Tracking cellular history with protein ticker tape By Brian O’Neill The Cohen...created a system for recording the transcriptional history of a cell. Lin et al. generated a fusion protein...ticker tape images could recover transcriptional histories, accurate to within 40 minutes. Figure ...

...of a TALE, along with strong nuclear localisation tags and repress transcription initiation when targeted... plasmids 47388 and 47389 include a Flag epitope tag and an SV40 NLS at the N terminus, a 152-residue ...the repeat domain, and C-terminal SV40 NLS and HA tags. Plasmid 47389 also contains the VP64 domain (four... has successfully been employed for deletion mutagenesis and gene targeting in human and mouse pluripotent... was recently shown that TALE fusions with the histone demethylase LSD1 (Lysine specific demethylase) ...with the FLASH method can decrease the amount of Histone H3 lysine 4 (H3K4) di-methylation at the target...

...Petri dish (VWR, 11019-552) 4 MaxiPreps (Qiagen HiSpeed Max, Catalog #12663) Tips (1000 µL, 200 µL, 10 ...Caution Electroporation involves the use of high voltages, please use caution when activating pulse and ...down at 30 ℃ overnight. Critical Ensure at this stage that no unabsorbed media drips onto the lid. Let... Maxiprep. Purify plasmid DNA using the Qiagen HiSpeed Maxi Kit (one conical is its own Maxiprep). Critical...

...hello. Keep your cards handy and make sure your name tag is always visible. Some people learn names by seeing... them. It is OK to ask someone to see their name tag if you are one of those people. Meet people in sessions...or make one!) or find the conference's Twitter hashtag. Use social media to let others know you will attend...Conference Dinner - by Julio Peironcely on the Thesis Whisperer "Not Networking - Building Relationships for Success...

...the sdAb, or by using a secondary against a tag, such as His, incorporated in the sdAb. sdAbs can have ...location. Includes: Immunofluorescence (IF) Immunohistochemistry (IHC) Immunocytochemistry (ICC) Cell Sorting...in these assays. This method is suitable for DNA:histone modifier interactions, which have strong binding...methods such as immunofluorescence (IF), immunohistochemistry (IHC), and immunocytochemistry (ICC) use... they only have one protein of interest. Immunohistochemistry (IHC) and immunocytochemistry (ICC) IF can... can refer to both immunohistochemistry and immunocytochemistry when fluorescent signaling molecules (...

...into a vector with a large tag at the N-terminus, but you don’t have the tag already included in the vector...you can insert both the gene of interest and the tag sequences into the vector in one step without scars...In this blog post, I will go over some advantages, disadvantages, and examples of how scientists are using... common, and for good reason. They offer many advantages over the traditional restriction enzyme cloning...thereof) nor a serious time commitment. Another advantage is that this process makes it easy to generate...will be using. Some of these products may offer advantages that Gibson assembly does not such as increased... 1. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA 3rd, Smith HO. Enzymatic assembly of DNA molecules...

... Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs...Available modifications include: histone acetylation by p300 histone demethylation by LSD1 cytosine methylation...for targeted epigenetic modification, including histone acetylation/demethylation, and cytosine methylation...

... lab of Arlene Sharpe at Harvard Medical School. His projects include investigating negative regulators..., largely depends on cell type and laser voltage. The voltage describes an electric potential that can... the cell or on its surface. Bear in mind, the voltage setting of the laser impacts the magnitude of FSC...setting as needed. To figure out the appropriate voltages to observe your cells on a flow plot, you might...respectively. H, A, and W describe the shape of a histogram, which graphically represents the time and intensity...graph of photocurrent vs. time. The resulting histograms is described through maximum current (H), the...the signal strength. So, we need to adjust the voltage in the cytometer’s software in order to generate...

...NanoLuc® with either SNAP‐tag or HaloTag7 tag in which labeling of the self‐labeling tag with a fluorophore ...and Renilla luciferase are less-than-ideal protein tags due to their large size. NanoLuc® Luciferase is ...biosensor that utilizes Renilla luciferase and a Halo tag to assay mRNA decay in real time. pKC-4.04, pKC-4.06...Lentiviral expression of firefly luciferase with a V5 tag Kevin Janes 83281 pAAV-CAG-FLuc Firefly CAG AAV expression...Mammalian expression of Nanoluc with a N-terminal Myc tag Erich Wanker 124701 pLenti-PGK-Venus-Akaluc (neo)...Lentiviral expression of Nanoluc with a N-terminal Myc tag Erich Wanker 115352 pFL-SV40 Firefly SV40 Mammalian...systems. Each type of luciferase has advantages and disadvantages depending on the application. Firefly...

...collections: Tags and Other Markers : Antibodies targeting popular markers like tubulin or epitope tags like ...species. We currently assess western blot, immunohistochemistry, and immunocytochemistry. Addgene will continue...