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... is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with restriction...program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...biology98.3 (1975): 503-517. PubMed PMID: 1195397. 2. Chang, Judy C., and Yuet Wai Kan. "A sensitive new...searching for), then the probe will bind to the sample thereby labeling it. The label can then be visualized...strength of the signal in comparison to a control sample. Southern blotting applications range from confirming... MstII and southern blotting are conducted on a sample from an individual without sickle cell anemia, ... largely replaced by real-time PCR to answer the same experimental questions. Real-time PCR detects and...

...upstream of an opsin gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents...Deniz, et al. "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ...general anesthesia (Taylor et al., 2016). These very same neurons also play a central role in motivated behaviors.... Hirsch, Matthew L., Sonya J. Wolf, and R. J. Samulski. "Delivering transgenic DNA exceeding the carrying...

... intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W...SSC-A, so most people use that setting. Figure 2: As the cell passes through the laser beam and causes...scale in proportion (while H and W do not; see Figure 2), single cell events will show up in a straight line...When using flow cytometry to analyze your samples, it is necessary to set up a sequence of gates to be...the area under the resulting curve. By default, a sample on the flow cytometer (before you set the first...Practical example: lymphocytes in a mouse tumor sample You made it! So much about the theoretical background...gated out my lymphocyte population from a tumor sample. Many cell types can be found in the tumor microenvironment...

...dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately equally dense... 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 ...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...time.   Samples can run into empty neighboring lanes, so load your gel to ensure your samples and reference...protein-based buffer (FBS, milk, etc.) against the same amount of sample.  If you’re using chemical blockers, you...

...Brainbow mouse neural labeling system. Brainbow-1 and -2 have been further refined to create Brainbow 3.2. ...M-CFP expression; these deletions are defined using 2 loxP variants (only identical loxP sites can mediate... each fluorophore. Third, Brainbow-1 and Brainbow-2 each contained a “default” state; for example, Brainbow...processes not previously visible with Brainbow-1 and -2. The general structure of Brainbow-1.0 is retained...three loxP variants, but the principles remain the same. The first fluorescent protein in the construct,...

...AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV-9 and...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates... PubMed Central PMCID: PMC3152950. 3. Jacobson, Samuel G., et al. "Improvement and decline in vision with...

... 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...contain the donor and acceptor fluorophore on the same protein and are useful when a process affects the...

...research experiences to undergraduate students (1, 2), but educators face significant challenges in designing... by western blotting in other experiments (Figure 2).   Complementation reveals conservation of function...Biology Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology...fission yeast, Schizosaccharomyces pombe, have the same function as their counterparts in S. cerevisiae....

... of components, such as your CRISPR enzyme or Pol 2 promoter. The options available in each class, or ...overhang specific to the type of module it is (Pol 2 protomer, Cas protein, ect...). Whatever individual... (KO, CRISPRa, CRISPRi, editing)   Figure 2: Selecting module fragments on the Fragmid website...(GG) cloning approach. Individual modules and a sample ligation are depicted. (Bottom) Schematic depicting...

...go anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes... the technology. This transition takes around 1 – 2 years. The product is the measure of success. Technology... into Sample6 as graduate students in Dr. Jim Collin’s Lab in 2009. From 2009 to today, Sample6 has changed...leadership, and grown to 25 members.  What is Sample6? Sample6 is an enrichment-free bacterial pathogen detection...current VP of Business Development & Operations of Sample6, for advice on how to take an idea developed during...etc. What project in the Collins lab inspired Sample6? The initial work focused on using bacteriophages... the focus of the application was changed, but Sample6 still leverages the core engineering and scientific...

...animal is given additional booster immunizations every 2-3 weeks to increase the antibody titer. The titer ...level of antibodies is reached - typically within 2-4 months. After the animal has the desired level of...recognize the same region, or epitope, of the target protein. These antibodies are also all the same isotype...heterogeneous mixture of many antibodies that recognize the same protein. In the immune system, antibodies are produced...peptide sequence derived from this protein. At the same time, an adjuvant such as KLH or Freund’s adjuvant...clones will still recognize your protein. For the same reason, they’re also useful in experiments where...experiment, you should either order several vials of the same lot of polyclonal antibody, or select a monoclonal...

... characteristics of select AAV serotypes in Table 2 from the paper. Delivery of AAV into the brain can...routes for neuroscience see Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific...intravenous injection. Even when injected into the same place in the brain, certain serotypes spread further...

...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...states in a sample; instead, it will suggest the predominant state of registers in a sample based on a ... use the same recognition site array on the register, and all three-input GRSMs use the same recognition...implement the same gene regulation program. We recommend testing multiple register designs for the same gene regulation...If an attB-attP pair is aligned (sites facing the same direction) on a register, then the result of their...

...groups, such as their vaccine production, SARS-CoV-2 research, or adenovirus packaging cell lines dataset...-Line Knowledge Resource. J Biomol Tech. 2018; 29(2): 25-38....hybridoma, etc.), as well as sex and age at time of sampling if available. STR profiling for human, mouse, ...

...represent the preexisting resistant subpopulation 2. Resistant cells among the separate replicates would ...medicine 21.5 (2015): 440-448. Pubmed PMID: 25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct...replicates would contain the same labels - i.e. they would always come from the same parent cells that represent...replicate groups, and treated these groups with the same drug regimen, they would find one of two outcomes...population (all barcodes can be amplified using the same sets of forward and reverse primers). The abundance...

...RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral ...than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...Figure 5: Mechanism of ATLASCre. VAMP2 = synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR...with TRV2 that encodes ISYmu1 and an ωRNA under the same promoter, they introduced heritable genome edits...

...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET...recombinant antibody in human postmortem brain samples. Results show positive staining in tissues from...

...(you generally don’t want more tag than protein), 2-3 tags is increasingly common. Tandem affinity purification...TEV cleavage site (more on that in a moment), and 2 ProtA IgG-binding domains. TAP has since come to encompass...found in this paper (available at Addgene).   Table 2: Protease recognition sites commonly used with tags...protein since the enzyme cleaves the SUMO tag in the same manner as it would have in the context of a cell...

... (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the presence... an introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! ...ending, we can still get invested in the journey? The same thing applies for science storytelling. You can ...

...demonstrated through living images created by the team (Fig. 2). These images are based on the concept of bacteriographs...spatially precise the activation is.   Figure 2: A bacteriograph of Michelangelo's "The Creation of...References Romano E, Baumschlager A, Akmeriç EB, Palanisamy N, Houmani M, Schmidt G, Öztürk MA, Ernst L, ...