We narrowed to 560 results for: cat.3

...end that is identical to an adjacent segment and a 3′ end that anneals to the target sequence. One strategy...reaction: T5 Exonuclease - creates single-strand DNA 3’ overhangs by chewing back from the DNA 5’ end. Complementary...) to the isothermal reaction mix. ET SSB protects 3’ ssDNA ends from the ssDNA-specific endonuclease activity...product, gel-purify DNA segments. Otherwise, PCR purification or even the raw PCR mix can work fine in an ...

...separated by size, (2) transferred to a membrane, and  (3) detected using antibodies.   Figure 1: Overview...post-translational modifications, quantify protein levels relative to standards, detect the cellular location of proteins...pull-down assays.  Westerns are divided into two categories, native and denaturing. In a native western, ...matrix, the blotting membrane, the gel, a copper cathode/iblot gel matrix, and filter paper. Pretty much...search, first pay attention to the validated applications listed on the vendor’s website and choose an...western and vice versa.  In addition to validated applications, pay close attention to the controls listed ...and incubation times. If the vendor includes publications that cited the antibody, consider reviewing ...

...scientific enterprise and not just impact. Cause #3: Poor experimental design It’s easy to get excited...someone has replicated it precisely. You can find a great discussion of different types of replication in Nosek...perfect story for a “high impact” publication can lead to the publication of rare flukes showing positive...that fall into this category include performing experiments with too few replicates leading to suspect ...these positive results will be published. The publication record will then show that the tested hypothesis...reproducible publishing practices. One example is the publication of peer reviews so we can see how and why papers... also means that you’re more likely to get a publication (for better or worse, the currency for success...

...in both mouse and zebrafish brains. Figure 3: PinkyCaMP and FRCaMPi in use. A) Confocal image of... and accessible education in gene editing and synthetic biology. Nature Communications, 15(1), 6563. https...cell mitochondrial transfer tracker Red calcium indicators PinkyCaMP and FRCaMPi Micropeptide killswitch...Workflow for cell-surface protein labeling and identification. Reproduced from Wu et al. (2025) under CC ...the Cre-tagged mitochondria, Cre is released, translocates to the nuclease, and excises the DsRed-STOP ...doi.org/10.1073/pnas.0510511103      Red calcium indicators PinkyCaMP and FRCaMPi By Mike Lacy Many popular...popular genetically encoded calcium indicators (GECIs) are available, but red GECIs have been limited compared...

...spend the next 6 years. Know yourself I rotated in 3 labs before choosing a graduate school lab. All the...honestly evaluate your own style of work and communication before making this important decision. I’ll ...Prepare by going sleuthing on the lab website, publication records, LinkedIn, ResearchGate or Twitter. Be...the appropriate first authorship position on a publication of her project and made another lab member first...– including failure to acknowledge good work, allocation of meaningless tasks, removal of responsibility...connections such as serving on Boards? Does the publication record show evidence of industry collaboration...skills guide Learn about careers in science communication Read posts on mentoring for scientists Resources...

...image-based figures for scientific publications. PLOS Biology, 19(3), e3001161. https://doi.org/10.1371...protein is (localization) and how much there is (quantification). But what if you’ve got several interesting...primary antibodies that have been validated in IF applications (find IHC validated or ICC validated antibodies...Immunocytochemistry Protocols for other antibody applications...

... 20, 2022 Workflow Timeline Day 1: Seed cells Day 3-4: Fix and label cells Equipment Pipette controller...a 24-well cell culture treated plate. Seed 5 x 10 3 HeLa cells per well. Allow the HeLa cells to grow ... min in 500 µL PBS on a rocking platform. Section 3: Labeling with antibody Block for 20 min at RT on ...Guide Western Blot Protocol Recombinant Antibody Purification Protocol Introduction Immunocytochemistry is...as methanol or acetone may be better for some applications. Remove the paraformaldehyde and follow your...

...0: Seed cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day 7 (am): Harvest cells Equipment...interest Triton X-100 Benzonase/DNAse I (Millipore 71205-3) 40% Polyethylene Glycol 8000 (PEG) + 0.5 M NaCl Cell... PBS and add 35 mL of 0.05% Trypsin/EDTA. Wait ~2-3 minutes for cells to detach. Gently tap the sides ...stir slowly at 4 °C for 1 h, then keep at 4 °C for 3 h without stirring to allow full precipitation. Precipitation... at 4 °C if needed. Transfer the entire sample to 3 x 500 mL conical bottles and centrifuge at 3900 rpm... of sonication to avoid overheating of the sample. Mix well between rounds of sonication. Sonicate until...container pH meter Stir plate Magnetic stir bar Sonicator Ear protection Vortex Reagents Adherent HEK293T...

...Originally published May 3, 2017 and last updated Sep 24, 2020 This post was contributed by guest blogger...the optimal location. It is also important to have good information on the exact location of the TSS. ...appreciable levels of modification. Whole-genome sequencing of cells modified by CRISPR indicates that the consequences...specific base edit, or modulation of gene expression. Location and sequence are important considerations for ... gRNAs. For indels, it's not so important what location in the gene you target, but it is important that...fewer sequences to choose from). Finally, for HDR, location is much more important because you have to target...target site is far more constrained by the desired location of edit; efficiency decreases dramatically when...

...and can exhibit recombination levels as high as 1-3%. In light of this finding, we now include guidance...promoter, such as the truncated tyrosine hydroxylase (TH) promoter for catecholamine neurons or glial fibrillary... Flp), to either flip or excise a gene that is located between them. Early recombinase-dependent vector...expression profile at multiple steps from plasmid amplification in bacteria to transgene expression in vivo.... the level of spontaneous recombination. This indicates that the amount of leaky expression can largely...are typically employed during viral plasmid amplification (Fischer, KB. 2019). This means that all DIO...of spontaneous reversion during AAV plasmid amplification. On the left homologous pairs of scrambled cre-recombinase...

...spectra Example: Jaws Archaerhodopsins Archaerhodopsin-3 (Arch) from Halorubrum sodomense is also commonly ...with single-spike temporal resolution. PloS One, 2 (3), e299. https://doi.org/10.1371/journal.pone.0000299... of distinct neural populations. Nat Methods, 11 (3), 338–346. https://doi.org/10.1038/nmeth.2836 PMID...of neural activity. Cold Spring Harb Protoc, 2011 (3), top102. https://doi.org/10.1101/pdb.top102 PMID:...Channelrhodopsins: cation channels from Chlamydomonas reinhardtii ChR2 Widely used light-gated cation channel from...light of the correct wavelength, the pore opens, cations flow into the cell (yellow dots), and the neuron...more information on biosensors such as calcium indicators or voltage sensors, visit our Biosensors Plasmid...

...about roles in this field, most jobs needed at least 3 months experience working in a developing tropical...at the Institute of Cancer Research in Sutton, located on the edge of London to gain relevant experience...recession. As each advertised job received many applications, employers were requesting that only scientists...commercial company. In my particular situation, communication between departments was sometimes difficult ... order to have meetings between teams. This communication is vital because otherwise I’d lose touch with...

...nanobodies. Nature Structural & Molecular Biology, 25(3), 289–296. https://doi.org/10.1038/s41594-018-0028...equipment on a lab bench. The best part? Less complicated math. Bioengineering is a field that has been... do we like it)? Antibody engineering is the modification of antibody sequences and structures to affect...extensively characterized and validated, with the application data from IPI and other labs easily viewed in...2021). Directed Evolution: Methodologies and applications. Chemical Reviews, 121(20), 12384–12444. https...

...Timeline Day 1: Seed cells Day 2: Transfect cells Day 3-6: Feed cells Day 7: Harvest antibody Equipment Biosafety...tube and vortex with three 1 s pulses. Incubate for 3 min at room temperature. Transfer the flask of HEK293...to mix. Return the flask to the incubator. Section 3: BCD Feed and valproic acid supplementation During...General Transfection Protocol Recombinant Antibody Purification Protocol Introduction Transfections allow for...antibody can be purified for use in a variety of applications. Sharing speeds science. We believe that sharing...antibody-expressing plasmids containing the SV40 origin of replication use a HEK293 line stably expressing the SV40 ... containing the Epstein-Barr virus origin of replication use a HEK293 line stable expressing Epstein Barr...

...anneal to the regions upstream (5’) and downstream (3’) of the DNA segment to be amplified. When these reagents... increase the time of the denaturing. Your 5’ and 3’ primers should be designed to have similar melting...Reverse Primers: Hybridize and are complementary to the 3’ ends of the sense and anti-sense strands of the template...adequately. Divalent cations such as Mg 2+ and Mn 2+ stabilize the buffer solution. These cations can also be ...used for PCR-mediated DNA mutagenesis. A higher cation concentration increases the error rate of the DNA... provide a suitable environment for the DNA amplification reaction. Reference Page Top Index...

...plasmid fingerprinting", where you cut the plasmid into 3-8 pieces such that all (or most) fragments are small...insert, but significantly off center (ideally around 1/3 of the way from one end), and also cuts in the backbone... . The pattern of the fragments on the gel can indicate if the plasmid contains the expected size insert...before use of more expensive forms of plasmid verification, such as DNA sequencing . In the example above...side of the insert, you can get a very obvious verification of the orientation so long as the expected products...

...Science 6.1 (2015): 50-69. Royal Society of Chemistry. 3. Ermolenko, Dmitri N., et al. "Observation of intersubunit...cellular impediments. FRET has been used in many applications, including drug discovery by big pharma (i.e... acceptor doesn’t need to go to the same exact location as the donor. As long as some of the acceptor ...C-terminus of your target protein, but even these modifications can affect the structure or function of certain... close to the donor, faulty delivery to other locations in the cell are not a problem, as long as they...The poly-histidine tag often used for protein purification is a reliable FRET acceptor labeling site when... way as NTA-sepharose beads used for protein purification: the NTA binds to the His tag on your protein...

... dynamics in mammalian cells. Biophysics Reports, 3(4–6), 64–72. https://doi.org/10.1007/s41048-017-0037...emission wavelengths, which are pivotal in their application in biotechnology (Grimm & Lavis, 2022). These... substrates/peptides and antibodies. Example applications of this include coupling to an antibody, whereby...whereby excitation of blue light will indicate where FITC is bound through green light emission. Key considerations...typically bind to. Stability of fluorescence: For applications that involve ranges of pH, and significant lengths...light, is another factor to consider. If your application requires longer periods of exposure, you’ll want...degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic nature. The potency and efficacy...

...would fail Addgene QC and would not be used. Section 3: Image Analysis Use ImageJ (Link opens in a new window...the curves for the protein bands as follows: Figure 3 Select Analyze . Select Gels . Select Plot Lanes ....Sample Peak 1 (contaminant) Peak 2 (contaminant) Peak 3 (HC) Peak 4 (LC) Total Area HC + LC Area Purity AR0018... Recombinant Antibodies Recombinant Antibody Purification Protocol Introduction This protocol describes...mobility of the bands (sample AR0016 in Figure 1) indicates that the samples may not have been processed correctly...ImageJ. Select File . Select Open . Choose the location of the file to open. Change the image type to ...select the Chart Type drop down menu and select Scatter chart. Change the data range and select the cells...

...chain-termination sequencing or dideoxy sequencing (Figure 3). Sanger sequencing relies on the random incorporation... efficient and reliable sequencing method. Figure 3: Summary of Sanger sequencing. Created with BioRender.com.... Uracil replaces thymine in RNA molecules. Every 3 nucleotides (codons) in a DNA sequence encodes for...Origin of Replication ( ori ) DNA sequence which directs the initiation of plasmid replication in bacteria... not located elsewhere in the plasmid backbone. In expression plasmids, the MCS is often located downstream...advantage of the process of DNA replication in vitro. During replication, the DNA helix is unwound by a... Educational Resources Molecular Biology Reference Molecular...