...digital PCR. Human Gene Therapy Methods, 25(2), Article 2. https://doi.org/10.1089/hgtb.2013.131 Wang,...(see maps section for images).     Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre...Digital PCR Method. Human Gene Therapy Methods, 29(2), 96–103. https://doi.org/10.1089/hgtb.2017.198   ...Samples of viral preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver...

...is excised from the donor vector.   Figure 2: The Gateway system adopts phage integration into ...our website to search for your favorite gene! STEP 2: Generate an Expression Clone When making the expression...expression construct in only 1 day, as opposed to 2+ days with traditional restriction and ligation cloning...Clon Transgen 4:138. doi:10.4172/2168-9849.1000138 2. Hartley JL. Use of the Gateway System for Protein...sequence of interest, you can move this DNA fragment across any expression system in just one recombination...

...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor... quantification. A consistent 5% positive error across the plate is fine; a 5% positive error on one side...until the multiplexed and individual values match across the board. If the samples or sample conditions ...

...protein expression across gram negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https:...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...transformation processes, and can be used by scientists across disciplines.  What confers broad host range in ...

...of other Cas proteins found in the so-called class 2 CRISPR-Cas systems. There are 24 variants of the cas14...the presence of infectious organisms (ex: SARS-CoV-2) and genetic mutations. By providing a gRNA specific...and intergenic regions containing the putative tracrRNA into a plasmid and expressed it in an E. coli ...

...typically, normal (bell curve) or skewed (see Fig. 2). Assumptions – characteristics of a data set; ...levels of variance among data.     Fig. 2: Examples of data distribution profiles.   Parametric... positive is much higher than if you only look at 2 or 3 pairwise comparisons. Similar to family-wise ...statistical facets of reproducibility. Adv Physiol Educ. 40(2): 248-252 (2016). 0.1152/advan.00042.2016 Finch, W...Simulation Study. Front. Appl. Math. Stat. Volume 2 (2016). doi.org/10.3389/fams.2016.00002  ... W. Holmes. Comparison of Multivariate Means across Groups with Ordinal Dependent Variables: A Monte Carlo...

...are many docking sites available for integration. 2. How soon would you like your flies ready for use?...beautiful immunofluorescent images like in this paper (2). Metabolic assays such as Seahorse can be employed...PMID: 16547094. PubMed Central PMCID: PMC1526508. 2. H. Jiang,  et al. Cytokine/Jak/Stat signaling mediates...because the transgenes are found in the same locus across fly lines (i.e. there won’t be positioning effects...

...required to make a plasmid. In 2023 (and at closer to 2 million plasmids shared than not!), we think it’s ...time does it take to make a plasmid? Figure 2: Survey results from a Twitter poll run in 2023, n...go wrong, replace them for free. Plasmids ship out 2-3 days after ordering, making it a toss-up as to whether...records and an organized lab saves time and money across the board. Consider alternative cloning strategies...

... will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models reinforcement...spend most of its times in the opposite chamber) [2]. Laser activation is usually controlled automatically...two different ways 1) your hypothesis is wrong or 2) there is some problem with the technical implementation...PMID: 21716290. Pubmed Central PMCID: PMC3775282. 2. McDevitt, Ross A., et al. "Serotonergic versus nonserotonergic...examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place preference...

...is funded have been well documented over the past 2 years (Alberts et al, PNAS 2014; Taylor, eLife, 2016...PMID: 24733905. Pubmed Central PMCID: PMC4000813. 2. Taylor, Justin Q., et al. “Avoiding a lost generation... Several Future of Research (FoR) symposia held across the USA during this time have revealed that young...Standards Act (FLSA) which mandates that all workers across the country either be paid more than $47,476 or...filed for union elections in only 10 departments across the university. Collective bargaining may provide...unique needs of students from different departments across the university and alterations to the student-university...

...that increase cancelled. Rounding out the remainder, 2% of postdocs who weren’t going to get a salary increase...with their plans, and 17 have cancelled their plans. 2 of those raising - the University of Illinois and ...$50,000 - higher than the level being raised to now - across a wide consensus of academic stakeholders. However...information as of December 1st. Salary inconsistencies across geographical regions When we originally started...when all others in Boston (some of them literally across the street from these institutions) are raising...

...proteoglycan (HSPG)3 AAV receptor (not strictly required)2 Human fibroblast growth factor receptor 1 (FGFR1... AAV2, so they are formally called rAAV2/1, rAAV2/2, and so on, although we often use abbreviated nomenclature...suggest these serotypes as a good place to start. Table 2: AAV tissue tropism for mice in vivo. To target... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PloS One, 8(9),...Retrograde Access to Projection Neurons. Neuron, 92(2), 372–382. https://doi.org/10.1016/j.neuron.2016.09.021...variants enabling potent muscle-directed gene delivery across species. Cell, 184(19), 4919-4938.e22. https://...

...and scholarships in different subject areas!    2. Resume and cover letter know-how  Tailor your resume...4. Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch... classroom cannot offer. Furthermore, companies across most industries tend to hire their former interns...student should use these guidelines consistently across a number of years as you apply for internships,...

...retinal cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved...would express Cre-mCherry and GFP.   Figure 2: Synthetic promoters can be paired for targeted expression...1345-1356. https://doi.org/10.1038/s41593-019-0431-2  Neff, E.P. An AAV library for retinal cell expression...retinas. By comparing the performance of promoters across these three species, the Roska lab concluded that...

... fluorophore excitation by absorption of a photon 2. Energy transfer from the excited donor directly to...PMID: 24194740. PubMed Central PMCID: PMC3810607. 2. Bajar, Bryce T., et al. "A Guide to Fluorescent Protein...PubMed PMID: 10500161. PubMed Central PMCID: PMC18018. 2. Miyawaki, Atsushi, et al. "Fluorescent indicators...fluorescent proteins, has led to the alternative acronym Fluorescence Resonance Energy Transfer. Unlike ...

...super-resolution or single-molecule techniques.   Figure 2: Two-color super-resolution imaging of ER and mitochondria...stimulated emission depletion (STED) nanoscopy. Scale bar, 2 μm. Inset shows comparison with standard confocal ...enables applications like multi-color imaging (Figure 2). If you’re working with purified proteins in vitro...Labeling in Living Cells. Chemistry & Biology, 15(2), 128–136. https://doi.org/10.1016/j.chembiol.2008.01.007...conjugates: A flexible chemical tag. Nature Methods, 2(4), Article 4. https://doi.org/10.1038/nmeth749 Strebinger..., Frangieh, C. J., Friedrich, M. J., Faure, G., Macrae, R. K., & Zhang, F. (2023). Cell type-specific ...

... anniversary! We've done so much in the past two (2!) decades that it's hard to summarize, but we wanted... reagents from 5,821 labs in 1,249 institutions across 55 countries.  Our first request was on September...materials to 254,034 scientists in 8,900 institutions across 111 countries. Just eight days into 2024, we've...

...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...to understanding how nuclear organization varies across the life of a cell, or how organization may be ...

...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...discovery of CRISPR/Cas systems, gene activation across multiple loci was an arduous process. When using...options to choose from when looking to activate genes across diverse cell lines.   dCas9-VP64 Description CRISPR... higher activation levels than SAM in some loci across Drosophila cell lines (Sajwan et al., 2019). SPH...

... of permeabilization and membrane integrity (Fig. 2). For example, saponin interacts with cholesterol ... at too high a concentration.      Figure 2: Examples of different permeabilization strategies...used cross-linker for IF. However, you might come across a couple other terms that can cause some confusion... just formaldehyde. Formalin: You may also come across formalin, which refers to a saturated solution ... cell membranes as they are not able to diffuse across natively. Permeabilization essentially punches ...