... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ... issues that can arise in this step: protein degradation, incomplete lysing/insoluble proteins, and protein...protein aggregation. Protein degradation, which would result in low protein yield, can be reduced by adding...

...neoblast cells and the gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating... characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific mRNA sequence...223). Link: http://abt.ucpress.edu/content/79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver...triggers a cascade of events that results in the degradation of the target mRNA and silences its expression...

...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone... on human artificial chromosomes. Montreal, QC, Canada 45.5017°N 73.5673°W – Vanessa Sung Hundreds of...de-funding of crucial environmental research programs, Canada has been here before.  These attacks on science...Research Centre at McGill University in Montreal, Canada.  Outside the lab, she dabbles in science policy...Intellectual Ventures Laboratory. Vancouver, BC Canada 49.2827°N 123.1207°W – Karissa Milbury I attended...

...resolved loci with physical map distances of 75 and 2 Mbp, with the calculated fluorescent distances correlating...established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that they could evaluate the...activation/repression. dCas9 has also previously been adapted to fluorescently label a sequence matching its ...

...literature? Very often the data of interest are in 1 or 2 figures from one paper and then another figure in ...ReFigure by creating ReFigures and answering this 2 minute survey! You can also find ReFigure on Twitter...the figures of interest along with the article metadata, legend and user created notes on a “Pinterest...

... fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell Sorter (FACs) A machine...photostability of your fluorescent protein when using 2-photon microscopy as this hasn't been well reported...PMID: 18703742. PubMed Central PMCID: PMC2606057. 2. Chtanova, Tatyana, et al. "Dynamics of neutrophil...immune system. The techniques used here could be adapted to studying other systems in which there is large... revealing the cellular behaviors that mediate adaptive and innate immunity in diverse tissue environments... to better understand T cell initiation in the adaptive immune response. It was known that T cells were...scale when using photoactivatable FPs - their degradation within labelled cells limits the timespan available...

... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; ...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...different fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various...of plasmids required for each input/output system; 2) removing all chemically inducible promoters; 3) improving...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...accuators. The researchers took a FRET reporter and adapted it into a BRET reporter by using NanoLuc® Luciferase...

...certain states of interest. For example, in Figure 2 below we present a two-input, five-state GRSM register...linking RSM inputs to endogenous gene expression (2). Our paper (1) explains how to arrange attB-attP...Science. 353, aad8559 (2016). PubMed PMID: 27463678. 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...straight-forward, concise explanation of how to adapt our engineered devices, recombinase-based state ...not expressed in each state. To help researchers adapt GRSMs for their research, we provide MATLAB-based...

...channelrhodopsin-2, commonly used in optogenetics, opening up the possibility of adapting GFP lines for ... Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation...transcription activation domains. Both systems are easily adaptable to neuroscience applications, including optogenetics...

... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing...method for genomic insertion. PITCh can also be adapted for whole-plasmid integration if you’d like to ...from a given promoter, and the technique can be adapted to other transgenes. It’s important to note that...increased versatility, it would be advantageous to adapt PITCh to insert genes into AAVS1, the “safe harbor...

...activation is.   Figure 2: A bacteriograph of Michelangelo's "The Creation of Adam." This bacteriograph ...demonstrated through living images created by the team (Fig. 2). These images are based on the concept of bacteriographs...

...recognize Fab fragments from the same species and 2) these fragments will be more likely to bind (and ...epitopes recognized by the secondary.   Figure 2: There are a few common solutions to avoid background...-Mouse Immunohistochemical Staining Technique Adaptable to Biotin-Free Reagents, Immunofluorescence, and...

...experiment using smFPs and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures...similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays...PMID: 25915120. PubMed Central PMCID: PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization...smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance probes for light and ...

... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...functions more similarly to CRISPR/Cas9 systems adapted for eukaryotes in that Cas9-mediated cleavage induces...addition of a uracil DNA glycosylase inhibitor and degradation tag on the fusion protein to limit the activity...

...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...folding, post-translational modification, protein degradation, access to the nucleus in eukaryotic screens ... system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira S.K., Chou J....Stynen B., Van Dijck P., Tournu H. A CUG codon adapted two-hybrid system for the pathogenic fungus Candida...

...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation signal directs...longevity in eukaryotes and promote transcript degradation in prokaryotes.  Prokaryotic termination   Prokaryotic...important for stability of the mRNA, protection from degradation, and is integral to the nuclear export and translation...

...episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student podcast episode 2 "Essential...This is a free web-based tool for detecting the readability of your writing. It makes suggestions on how ...

...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor...disrupting ribosome-mRNA complexes. Recent work has adapted this technique to monitor local translation and...

...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241  ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695  Schmidhauser...contains structural elements that are versatile and adaptable. Variation in ori structure allows plasmids to...