We narrowed to 823 results for: lars

...Restriction enzyme cloning is the workhorse of molecular cloning; however, one of its biggest limitations...The best applications for dsDNA inserts include: large insertions or deletions, including selectable DNA... sure to read the associated papers for their particular uses - *note* not all linked strains contain ...is to use mini-λ, a defective non-replicating, circular piece of phage DNA, that when introduced into ...generation of bacteriophage, which is not a common molecular biology technique. However, an advantage of this...Medicine at the University of Washington. She is particularly interested in science communication and in vitro...PubMed Central PMCID: PMC307639. 2. Daiguan Yu, Hilary M. Ellis, E-Chiang Lee, Nancy A. Jenkins, Neal ...

...) is conceptually similar to the microRNA precursor mentioned above and is a popular RNAi trigger in the...exquisitely and robustly self-organize at the cellular and multicellular scale.     References Bezanilla M, Perroud...is fair to say that CRISPR has revolutionized molecular biology and has enabled functional studies in ...micro RNAs (amiRNAs) are frequently used. Although similar, amiRNAs are engineered from endogenous precursors...silencing plants. Non-destructive methods, most popularly with a fluorescent reporter, directly couple silencing...Institute where he investigated how plants achieve polarized cell growth in the lab of Luis Vidali. For his...10.1242/dev.186452 Wilson RC, Doudna JA (2013) Molecular Mechanisms of RNA Interference. Annu Rev Biophys...

...is often the case with large genes. This is due to inefficient packaging of large inserts; a study by Kumar... While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges... deciding what packaging system is best for a particular experiment. The newer generation lentiviral packaging... guides) while a single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example...dishes or increasing the vessel size. Opting for a larger vessel, such as a 3 or 5-layer flask will require...calculating the ratio of the surface area of the larger vessel to the 10 cm2 dish and adjusting the cell...start. If you don’t anticipate that you will need large volumes of lentivirus routinely, you may opt for...

...vertical.  Large central box. Another commonly used and effective style is one employing a large central ... figures fit nicely into circular shapes rather than a square or rectangular boxes. Background and color...poster together, be aware that there are some particular styles recognized to work well. It is important...is written across the center of the poster in a large font. Hence, spectators can read the text from many...written and graphical information are inserted into larger circles or ovals, that are arranged in some logical...these colors so that your text is easily read. Similarly, be cognizant of audience members that are color-blind...

...improved CRISPRa, expanding its practicality and popularity in research (Tanenbaum et al., 2014, Konermann...Konermann et al., 2015, Chavez et al., 2015). Most popular methods for CRISPR activation CRISPR activation ...compared to other CRISPR activators, making it a popular method for gene activation experiments. In cases...exhibits activation levels comparable to other popular activation methods (VPR and SunTag) (Chavez et ...relies on antibody chains, which are relatively large and are not expressed consistently throughout cells...compared to the SAM system. Its activation levels are similar to that of SunTag. An advantage to this method ...While SAM, SunTag, and VPR have become the most popular methods for CRISPR activation, research in the ...

... and circular assemblies. While Golden Gate Assembly worked for all the assembly types, circular assemblies... 52-piece circular assembly of the T7 genome had lower efficiency than the 10-piece circular assembly,...rules the best way to design GGA overhang sets, particularly for high-complexity reactions? The researchers...outlined below, allow for researchers to select large sets (with overhangs that violate some of the traditional... use to identify fusion sequences for either a circular or linear assembly. If you aren’t constricted ...fairly easy cloning technique (which may be of particular interest to MoClo users!). If you’re interested...

...Find basic molecular biology protocols Find plasmid cloning protocols Check out our molecular biology reference...one-pot” reaction to form complex, multi-insert modular assemblies that enable biosynthetic pathway engineering...destination vector receives the assembly after similar Type IIS restriction enzyme cutting results in ...performed additional efficiency measurements using similarly sized Lambda amplicon inserts to control for any.... To compensate for this, researchers can plate larger volumes of the outgrowth containing the transformants...assembly. The fidelity data can be applied to derive similar high-fidelity overhangs for any Golden Gate assembly...England Biolabs, Inc.         Eric Cantor is a molecular biologist and a Development Group Leader overseeing...

... are particularly good models for building chromatin interaction networks since they are a large cluster...biotinylated by BirA. From here, the CAPTURE method is similar to other 3C methods: crosslinking captures chromatin...Additionally, CAPTURE 3C-based sequencing requires a large number of cells (~5x107). Together, these two requirements...well-characterized beta globin locus. The two methods had similar rates of specificity for the DNA sequence targeted...interactions identified with the multiplexed CAPTURE 2.0 largely replicated interactions previously identified with..., like those that occur during development and cellular differentiation. Using a cell line model, the ... for spatial genome organization deciphering. Molecular Cytogenetics, 11. https://doi.org/10.1186/s13039...

...single base pairs, to chromosomal inversions and large deletions. Many of these methods rely on Cas9 or...Although seemingly ubiquitous in nature, they are largely underutilized in research settings — mostly a consequence...is not as well understood but likely occurs in similar regions. This combination of chewing up one stand...stand and cutting the template strand causes large deletions of up to hundreds or even thousands of base...proteins naturally bound to the DNA. Cas3 or Cas9? Large genomic deletions can be achieved using Cas9, but...but Cas3 may be a better option. To get large deletions using Cas9, two or more gRNAs must be used along...Cascade-Cas3 complex in nucleotide unwinding steps. Molecular Cell, 70(3), 385-394.e3. https://doi.org/10.1016...

...at the molecular level, at the cellular level there is a universal theme - uncontrolled cellular proliferation... no other disease receives as many billions of dollars in funding or as much attention collectively from...causes of this uncontrolled proliferation, in particular the roles of proto-oncogenes, tumor suppressors...the accelerator of a vehicle being stuck down. Similarly, a mutation that turns a tumor suppressor “off...universally-dividing behavior of cancer cells, attacking the cellular machinery necessary for division. This approach...research has focused on picking apart the complex cellular pathways involved in the development of different...

...and Biosensors: Throwing color lights on molecular and cellular processes” Curr. Protein. Pept. Sci. 9(...discovery, the family of FPs just keeps getting larger with hundreds of variants available. Read on to.... Therefore, these can be used as “timers” for cellular processes following their activation. The four... called Slow-FT, Medium-FT, Fast-FT, and mK-GO. Large Stokes Shift (LSS): Stokes shift (named after George...split FP. This technique is also is also called Bimolecular Fluorescence Complementation (BiFC). 8 points...The maturation of the chromophore on many FPs (particularly those derived from GFP) requires oxygen. Therefore...interested in everything related to gene expression, particularly at the RNA level. He maintains the greenfluorescentblog.wordpress.com...

... activated at a similar spectrum as HR and also results in membrane hyperpolarization but unlike HR, this...Simon! Derek P. Simon received his PhD in Cellular and Molecular Biology from the University of Michigan...neuron in a specific brain region can result in a particular neurotransmitter release or post-synaptic current...the kinetics of the opsin variant, and other particularities that best suit your specific experiment. Consult...cation channel, activation of which results in depolarization and subsequent firing of an action potential...results in efflux of chloride ions, subsequent hyperpolarization, and inhibition of action potential generation...crossed with an appropriate mouse line expressing a particular cre, viral expression of your opsin is no longer...

...the most popular and useful viral vectors in the lab. Advantages of lentivirus include a large genetic ...genetic cargo (8 kb vs 4.5 kb), and they remain popular for targeting and tracing cells in the brain (Figure...vectors in the lab Lentiviral vectors are incredibly popular in the lab - the Trono lab’s lentiviral packaging...the clinic Since lentiviral vectors can deliver a large amount of DNA (~8 kb) with a relatively low immune...lentiviral vectors for applications that require the large genetic capacity of a lentiviral vector but for ...PubMed PMID: 23322669. 6. Genetic treatment of a molecular disorder: gene therapy approaches to sickle cell...Dorazio R, Mulder GJ, Polack A, Bank A, Soulier J, Larghero J, Kabbara N, Dalle B, Gourmel B, Socie G, Chrétien...

...minigenomes are incapable of producing infectious virus (similar to lentiviral vectors) because they do not encode...components of the Zaire ebola virus minigenome system Similar to other RNA viruses, EBOV brings its own RNA-dependent...which is formed by a complex of two proteins: the large (L) catalytic protein and its cofactor VP35. VP30...transcribed (and the viral mRNAs are translated) by cellular machinery. NP encapsidates the negative sense,...necessary control elements for each would be relatively large. Cloning the viral genes in this plasmid would be...polyadenylate the mRNA. These mRNAs are translated by cellular host machinery (as viral mRNAs would be in the...processes in extreme detail. The mechanism of a particular observation can be easily studied as each component...

...the entire proteome of a particular cell type, or a subset based on a cellular compartment or growth stage...Obrdlik, P., El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, H., Kamuzinzi... and are still used extensively to screen for molecular interactions in the cell, including protein-protein...activities (1). Researchers also discovered the modular format of some transcriptional activators: that... yeast two-hybrid (Y2H) system as a robust and popular lab tool. In the classic two hybrid system (also...two-hybrid systems are a powerful tool for screening molecular interactions, accuracy and reproducibility are...

...one (admittedly large) class of common fusion protein tags. Biochemists and molecular biologists who need...challenges, researchers have amassed a considerable molecular tool box of tags and fusion proteins to aid in... interest to overexpress a protein. Energy and cellular resources are being spent to make something the...small sequence of amino acids, is basically a molecular leash for your protein. If you’re working with...; has inherent enterokinase cleavage site GST Large Protein 26 Purification and Stability Good for purification..., flanked by hexahistidine motifs (6xHis) MBP Large Protein  40 Solubility and Purification Can improve...Solubility and Affinity Abundance of charged and polar residues improves solubility; good for antibody-...

.... However, most lesioning methods destroy areas larger than intended and may unintentionally remove connecting...an animal’s movement in an open field box or a circular corridor apparatus using a camera or tracking ... psychiatric diseases such as schizophrenia or bipolar disorder. In both diseases, animals have reduced...time in the open arm and were less risk averse. Similar to the elevated plus maze, the light-dark box is...protected) area. When measuring the time spent in a particular part of the apparatus, it turned out that the...downregulating expression of miR124a achieved a similar improvement in the forced swim test.  Another brain...this further and showed that the activity of a particular subset of excitatory neurons (Drd1) in mPFC is...

... SPH mice CRISPRa has largely been limited to in vitro settings due to the large size of Cas9 activators...activators, which largely precludes delivery by adeno-associated vectors (AAV). This issue was recently circumvented...delivered to the CNS by lentivirus, which has a larger packaging capacity than AAV. Such an approach was...neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church Lab. Neuron-specific...microglia are a notable exception) necessitates such molecular gymnastics for sensitive applications such as ...screening. The plasmids used by the Qi Lab for this particular application can be found here.  Why CRISPRa? ...well-suited, including studying mRNA dosing effects, cellular reprogramming, and disease modeling. Further, ...

...model organism blog posts: Five Popular Model Organisms, Part 1 Five Popular Model Organisms, Part 2 ...In this heyday of molecular biology, many scientists do a lot of DNA work but never get to actually manipulate...materials sharing in their dissemination plans and in particular, that they all share the plasmids created under...which have been used to treat heart disease. Caterpillars that eat milkweed generate cardenolides that... poisonous) for bird predators - birds eat a caterpillar once, vomit, and never do it again. Cichlids ...Crepidula atrasolea juvenile, light microscope with polarizing filter. Development is direct in this species... fascinating social behavior. For example, two similar species have very different social behavior. One...

...important regulator of embryonic development The largest conference room at Addgene is named Sonic Hedgehog...segmentation during embryogenesis. Typically, regularly spaced denticles (bristles) outline the segments...contain SHH, you can find them all here! The most popular SHH plasmid however was deposited by none other...transposons as a non-viral system to integrate particular genes into mammalian genomes and easily disrupt...Ivics et al 1997. Spaghetti Squash, a regulator of cellular contraction You may think that a gene called "...the fly actomyosin cytoskeleton that regulates cellular contraction. This network is composed of actin...find the exact origin of this gene name but the larval gut of bagpipe mutants vaguely resemble a bagpipe...