... 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...

...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...

... 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...

...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...

...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 ...

...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...

...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...

... 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...

... Research, Ludwig-Maximilians-University Munich. His lab develops iPSC-based models of neurodegenerative...introduction of an intended mutation while taking advantage of the improved efficiency provided by blocking...

...FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate protein localization...fusion, etc). For example, by engineering PA-GFP tagged to a mitochondrial matrix protein (mito-PAGFP) ...Markus, and Wei‐Lih Lee. "Photoactivatable GFP tagging cassettes for protein‐tracking studies in the budding...an FP-chimeric protein may be at any one of many stages of synthesis and degradation. For this reason it...reversible photoactivation - and have enabled sophisticated imaging techniques.     Irreversible photoactivation...victoria) GFP (wtGFP) by mutating Threonine 203 to Histidine (T203H) (1). wtGFP normally contains a mixed population...PA-FPs by performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry...

...of CRISPR in 2013 (Cong et al., 2013)! Haoyi got his first exposure to CRISPR when he used this new technology... embryos as Cas9 from S. pyogenes and has the advantage of being smaller (Zhang et al., 2016). For editing...replacement of the murine sequence with a human gene. Advantages of using CRISPR for mouse genome engineering ...the efficiency discussed above extends 4 major advantages of CRISPR mouse editing when compared to more...Post for Tips on CRISPR Gene Editing in Mice Disadvantages of using CRISPR for generating mouse models ...

...blogger Jacob Lazarus from Harvard. Jake received his MD and a PhD in Cell and Molecular Biology from the...switched over to pTOX. However, we feel it has clear advantages, and are hopeful that continued development of...

...Fluorescent Protein Should I Use? However, fusion proteins tag your protein of interest with a fluorescent reporter...protein. One downside of fusion proteins is that the tag can alter the structure of the protein. In some cases...timing of expression can also be affected by the tag.  Protein expression, measured using antibody conjugated..., determining cell health, analyzing cell cycle stages, and detecting specific populations of cells in...system converts the fluorescence intensity to a voltage pulse, called an event, and assigns each event ... then gate the B cell population and create a histogram of CD19-containing cells versus total cell number...

... still infect the researcher and insert into her/his genome - this is a biosafety risk Infects dividing...to infect your cells of interest.   Advantages Disadvantages Can carry large transgenes (>8 kb) Potential... cells, it was obvious that its safety was a disadvantage particularly for clinical development. In addition...Efficient gene transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human...

...likely to lead to scores. That player doesn’t trust his teammates; the teammates don’t trust him. This team...Meselson-Stahl, Hershey-Chase...Who was I to dispute history? And boy did I make the right decision. If I’d ...the first place? Let’s think about some of the advantages of team-based work: Nobody knows everything anymore...Jefferson dined alone.” There was a time in human history when an intelligent, motivated person really could...

... lab of Arlene Sharpe at Harvard Medical School. His projects include investigating negative regulators...itself, like laser type and power, acquisition voltage, the concentration of the fluorophore, and whether...bd-spectrum-viewer.   But what is the real-life advantage of a brighter fluorophore? Strong brightness makes...

... vectors for experiments encoding a fluorescent-tagged molecular tool.   Find pAAV-FLEX-tdTomato, pAAV-GFAP104...variant in AAV1 at Addgene! Archon1, a voltage sensor AAV Historically, electrical activity in the brain was...measured with implanted electrodes. The Archon1 voltage reporter allows a way to monitor neuronal electrical...Archon1-KGC-GFP-ER2]) of this membrane-embedding voltage detector. Find Archon1 and its cre-dependent version...pAAV-hSyn-EGFP *Also coming in our new 20 ul trial size! Voltage indicators Plasmid Serotype Name 119036  AAV1...

...proteins in E. coli (Zhu et al., 2022). It uses the TAG codon for the phosphoserine and, because it uses ...technology or as a therapeutic.  A (very) brief history GCE came onto the synthetic biology scene in 1989...essentially those needed to make site-directed mutagenesis, many researchers new to GCE aren’t able to successfully... V. W. (2021). Genetic Code Expansion: A Brief History and Perspective. Biochemistry, 60(46), 3455–3469...