We narrowed to 780 results for: scr
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TypeBlog Post...looking at your resume while they wrote the job description. You check off all the boxes - you have the desired... gaps in scientists' transferrable skills. In describing the limitations of the study, the authors noted...over email. Examine your current and past job descriptions. Scan through all the things for which you are...
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CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog Post...Cas13a as a diagnostic tool, as discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a ...Recombinase Polymerase Amplification (RPA) or Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides... the collateral RNA cleavage response is well described in bacterial cells, LwaCas13a does not display... -
How to Write a Scientific Review Article
TypeBlog Post...we probably exchanged dozens of drafts of the manuscript. This is the time to transform the document into...Resources on the Addgene Blog Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing ...Organizing Tips for Improving Your Next Manuscript 6 Tips for Grant Writing Resources on Addgene.org Check out... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog Post...amplification) or reverse transcriptase (RT)-RPA, respectively. RPA is coupled with T7 transcription to convert amplified...2018) CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science 360... -
Supporting Reproducibility with a Connected ELN
TypeBlog Post...used in research, and data repositories. Below I describe how each of these kinds of connectivity facilitates...deposits will be made. The Dataverse integration is described in this video and the Figshare integration in ...from RSpace documents to external data sources (described above) further enhances the value the repository... -
Bacterial Toxin-antitoxin Systems as Molecular Biology Tools
TypeBlog Post...restriction endonucleases in plasmid addition was first described by studies of EcoRI and Bsp6I in Escherichia coli...systems. As with the other toxin-antitoxin systems described above, one part of the pair is more stable than...toxin-antitoxin system discovered. Unlike the systems just described, the hok/sok systems consist of three components... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog Post...heterologously by bacteria inside the body. So we screened gas vesicles gene clusters from a diverse set ...expression heterogeneity required single cell screening to isolate a clonal cell line capable of producing...improvement in expression. Stabilizing the gvpA transcript with WPRE reduced the number of copies of gvpA... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog Post...vector with two other independently regulated transcription units. Created with BioRender.com. Planning...contain plasmids with undesired ligations, so always screen multiple colonies after each transformation. This...beginners or those performing complex assemblies should screen a minimum of 8–10 colonies. You can check colonies... -
Changing Labor Laws Bring Increased Postdoc Wages
TypeBlog Post...expected for postdocs in many cases should be as described in the notice of the new NRSA levels and in the...very relieved not to be going through the urgent scramble of their peer institutions. The issues facing ..., and it is becoming ever clearer that outside scrutiny of science and the academic system is increasing... -
Deep Mutational Scanning with One Pot Saturation Mutagenesis
TypeBlog Post... The one-pot saturation mutagenesis technique described in Wrenbeck et al is a PCR-based approach for ...library that’s ready to be tested in a functional screen of your liking. This method is similar to site-directed... screen... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog Post...strong transcriptional terminator BBa_B1002 (from IGEM) upstream of the MCS to reduce transcriptional readthrough...worked too. Rhamnose induction relies on the transcriptional activator rhaS. To allow manipulation of bacteria... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog Post...same fluorescent markers, Brainbow 3.1 and 3.2 (described in the same publication) are unique because by...cell type. The Confetti mouse was first used to describe the self-renewal of cells in the intestinal crypt...previous Brainbow technologies were useful for discriminating neurons within only one field of view. Tetbow... -
Using Phosphoserine to Study Protein Phosphorylation
TypeBlog Post...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool...context-specific effects of UAG suppression, as described (6). To determine the stoichiometry of phospho-incorporated...phospho:non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid... -
CRISPR 101: Homology Directed Repair
TypeBlog Post...Easi-CRISPR in vitro transcribes RNA encoding your repair template, then uses reverse transcriptase to generate..., and ultraviolet light. In this post, we will describe the mechanism of homology directed repair, one... -
Hot Plasmids: Summer 2024
TypeBlog Post...methyltransferase — that can drive lasting transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration...versions using dCas9, zinc finger proteins, and transcription activator-like effectors (TALEs), with fifteen...Weissman, J. S. (2021). Genome-wide programmable transcriptional memory by CRISPR-based epigenome editing. Cell... -
PiggyBac-ing Through the Genome Editing Field
TypeBlog Post...applications, including transgenesis, gene-trap screens, and gene editing. How does the piggyBac™ system...effects, such as with signaling molecules or transcription factors, or when the gene is constitutively-..., since there is no selection, you will need to screen somewhere between one hundred and one thousand ... -
Antibodies 101: Validation
TypeBlog Post...Antibody-independent methods may include analysis of transcriptomic or proteomic data from several different tissues... feasible and (2) large-scale proteomic and transcriptomic datasets already exist. Example: You want...tissues. You know that there is a wealth of transcriptomic data from this sample type, so you use your... -
Special Delivery: Fluorophore Targeting for FRET Studies
TypeBlog Post... drug discovery by big pharma (i.e. HTRF and LanthaScreen) (Degorce et al., 2009), measuring intracellular...are critical in setting up a FRET-based assay or screen (the Addgene blog has an excellent primer on FRET...it all together The FRET donors and acceptors described above can be paired together to measure a wide... -
Antibodies 101: Designing Your First Flow Panel
TypeBlog Post...brightness of individual fluorophores. Table 1 describes the brightness of a few commonly used fluorophores...brightness levels on your smartphone or computer screen), and you should be able to properly label many...panel! Designing a Panel In this example, I will describe a basic panel (Table 2, Figure 5) for a hypothetical... -
Deep Dive: qPCR
TypeBlog Post... by abundant transcripts, leading to falsely lowered levels for less abundant transcripts. However, adding...can even be used to detect a single copy of a transcript. But this sensitivity can sometimes cause as ...