...other words, newborn poop) visible through their translucent bodies. This is the ideal stage to select for... transgenic flies...

...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9... the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987). The epitope (GKPIPNPLLGLDST) has...10.57733/addgene.l0ulu4. Hanke, T., Szawlowski, P., & Randall, R. E. (1992). Construction of solid matrix-antibody-antigen....org/10.1099/0022-1317-73-3-653. PMID 1372038. Randall, R. E., Young, D. F., Goswami, K. K., & Russell...functional properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024). They...

... RNAs inhibit plasmid replication.   Figure 1: The replication problem. Iterons are repeated ...actually does, and neither plasmid is maintained (Figure 1).  The partitioning problem Plasmid incompatibility...Agrobacterium tumefaciens. Genes & Genetic Systems 77:1–9 . https://doi.org/10.1266/ggs.77.1 Pubmed PMID: ... and inhibit its own transcription. As copy number increases, RepA transcription is repressed and replication...indirectly inhibit replication through inhibiting translation of replication machinery proteins, or directly... division. They don’t simply rely on chance (or random diffusion) to make this happen.  There are several...CBPs and NTPase to go around, the plasmids are randomly positioned, leading to plasmid loss (Diaz et al...

...sentences) What will be the focus of your first grant? (1-2 sentences) Simply stated, what are your...Full "Applying for a Faculty Position" eBook Here 1. Have a competitive CV You don't necessarily need ...that people would agree is important to understand. (1-2 sentences). For example: Distinguishing correctly...emphatically deny that I have a magic formula to guarantee a reader the faculty job of his/her dreams. There...currency of scientific success is publications and grants. 2. Write succinct and easy to understand documents...years. You will discuss the focus of your first grant, how you will overcome certain obstacles, alternative...It's not about you and how you will finally be granted the faculty position you have craved since grad...

...protein sequence of the predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is optimized ...several other constructs in plasmid form. Fig. 1: Summary of the approach and outcomes of the screen...voltage indicator: JEDI-2P  by: Brian O'Neill  The François St-Pierre lab and collaborators have successfully...photoactivation of HcKCR1 in mouse neurons. D) Membrane voltage traces of action potentials in a HcKCR1...

...screening for the best gRNA Figure 1: Validation of double strand break (DSB) mediated homology dependent...as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence indicates a more...and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target effects...repair (HR, homologous recombination; SSA, single strand annealing) took place and reconstituted the EGFP...site. (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences...efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP...fluorescence, the better that gRNA is at causing a double stranded break at the target site, allowing the two halves...

...derekpsimonphd.    References 1. Stuber, Garret D., et al. "Excitatory transmission from the amygdala to nucleus...animal will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models... can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem...examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place preference...(viral expression, efficiency of laser light transmission to the target neurons, appropriate stimulation...optogenetics is performed in this species. However, transgenic rat technology is improving and may one day reach... reach the sophistication of mouse transgenics. Everything that has been described is intended for the...

...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...on the Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of the...would the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem...restriction digest of plasmid DNA, and bacterial transformation. And more plasmid-related educational resources...

... are not required to initiate the reaction. Table 1 compares some general advantages and disadvantages...disadvantages of luminescence versus fluorescence.  Table 1: Properties of Luminescence versus Fluorescence   ...codon, which is then transfected into the desired cells. A hybrid mRNA transcript containing luciferase...luciferase, usually serves as an indicator of transcription within cells, where detection of the reporter...Higher Lower Background Lower Higher Post-translational Modification Not Required Required Photobleaching...regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these...as the most efficient means to disrupt the cell membrane and deliver the substrate; however, secreted luciferase...

...automated. Unlike gel electrophoresis, you only need 1-2 ul of sample and the run time is just a few minutes...quantified first before larger fragments. Figure 1: The quantification of a RNA ladder using capillary...This method involves measuring the absorbance/transmission of light through a liquid to determine the concentration...spectrophotometer measures these absorbances using UV-transparent cuvettes. First you measure the absorbance of...PicoGreen or SYBRGreen, are specific for double strand DNA (as compared to spectrophotometric methods ...However, the measurements are made in the visible range and can be read with a standard ELISA reader if ...

...References 1. Bressan RB, Dewari PS, Kalantzaki M, Gangoso E, Matjusaitis M, Garcia-Diaz C, Blin C, Grant V, ...the tag sequence flanked by homology arms (Figure 1). In the first step of optimization, we started off...form the ribonucleoprotein complex (csRNP, Figure 1). We use in-house purified Cas9 protein, which, in...standard immunocytochemistry (ICC) assays.  Figure 1: Three ingredients of epitope-tagging experiments....binding to the template strand (strand used by RNA polymerase II for transcription) are more effective for...complex. This complete transfection mix, i.e. csRNP plus ssODN, is then transfected into stem cells. Cells... were transfected in 96-well format using an Amaxa Shuttle device; 5 days after the transfection, the ...

...two weeks, we were able to obtain eight clones. We ran digested and undigested samples of the clones on ...intern to complete and present on.    Figure 1: Addgene's summer interns presenting their capstone...and I wouldn’t want it any other way. Toj Toj-Villagran is a rising senior in high school who just completed...

... the highly upvoted question.”If the Q&A session ran out of time, the speaker could continue to answer...participants could share their notes together. Figure 1: Can you draw Spongebob from these instructions without...

...nLightR (Kagiampaki et al., 2023).     Figure 1: Properties of nLightG and nLightR. A, Structural ...Article 6. https://doi.org/10.1038/s41592-022-01488-1 More resources on the Addgene blog Fluorescent proteins...generating variants with improved affinity and dynamic range of both nLightG and nLightR, and may soon have a...

...following three strategies might help!     Figure 1: Species on species staining can be a problem because...same host species as your secondary antibody since 1) your secondary should not recognize Fab fragments...References and Resources References Goodpaster T, Randolph-Habecker J (2013). A Flexible Mouse-On-Mouse Immunohistochemical...

...your budget, you should take the following steps: 1. Define your timeline Start with your thesis in mind...these decisions in the following order of importance: 1. First make decisions based on experimental timelines... several factors aren’t taken into consideration. 1. Always focus on the science When negotiating finances...will take to secure your first (R-level) research grant.  Set your timeline from there, then start identifying...techs’ financial books.  To find personnel salary ranges, go to online job information sites (like, indeed.com...

... by PCR has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million bp depending on...chance for mis-priming, so you can use a pretty wide range of annealing temperatures, but you may need to increase...

...the journal-specific guidelines.  Peer-review step 1: The first read At the first read of the paper, I ...group of Prof. Mark Tester, where she focuses on (1) salt-induced changes in root-to-shoot ratio in Arabidopsis... discipline and reduces the dissemination of unwarranted claims or unacceptable interpretations. A good...studying plant development and abiotic stress tolerance, I have been peer-reviewing journal submissions...

...sequences are inserted into the genome. Figure 1: Self-excising cassette schematic from Dickinson et...):1028-34. doi: 10.1038/nmeth.2641. Epub 2013 Sep 1. PubMed. Find plasmids from this publication at ...can generate a fluorescent protein fusion, transcriptional reporter, and loss-of-function allele in just...markers. Bob Goldstein’s lab previously used a transgene selection strategy for their C. elegans/CRISPR...

...those ideas and build from those.   Figure 1: Virtual lab meeting? No problem! You can still creatively... on it – so it is important to take breaks. After 1 hour (or another designated time) of meeting, someone... creativity are pivotal in writing publications/grants too. As described by several works (Wuchty et al...and do a quick break to chat. This can be done randomly or manually assigning which participants to group...