We narrowed to 842 results for: Eras

... are typically DNA preps, but we do distribute several of our most popular CRISPR pooled libraries and... A) Amplification, validation, and lentiviral generation of a pooled library. An aliquot of the pooled...experiments. Pooled library lentiviral preps are generally aliquoted to reduce the number of freeze-thaw ...libraries we amplify in house, but these are not general amplification protocols. Rather, each protocol ...amplified, you'll need to verify your library by next-generation sequencing (NGS). The representation of your ...our protocol on Lentivirus Production provides a general approach for producing lentivirus in Lenti-X 293T...you can transduce your cells.  Titering After generating your lentivirus, you’ll want to infect your cells...

... 101 blog, we reviewed the salient features of several popular strains of E. coli for DNA propagation....DE3) F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) General protein expression BL21 (DE3) pLysS* Chloramphenicol...ompT lon hsdSB (rB- mB-) gal dcm rne131 (DE3) General expression; not recommended for toxic proteins ...lon hsdSB (rB- mB-) gal dcm araB::T7RNAP-tetA General protein expression  BLR (DE3) Tetracycline RecA-deficient...miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein expression  m15 pREP4*, ** Kanamycin ...induction system. The DE3 lysogen expresses T7 RNA polymerase (RNAP) from the bacterial genome under control...overexpression? The strains described above should generate sufficient expression levels for most purposes...

... for monitoring how both the brain and visceral organs interact during various behaviors. Either tissue...very cool tool to advance our knowledge of brain-visceral organ signaling. The problem  Previously, there...emit light for optogenetic purposes, measure temperature, study electrophysiology, and deliver small molecules...molecules.   Fun fact: A 1 cm block of polymer can generate many meters of fibers!     Fig. 1 - ...for gut implantation. The fibers they ended up generating were 10-15 times more flexible than the brain... while brain tissue is relatively static). To operate both types of implanted fibers simultaneously, ...multiple channels and can also record tissue temperature proximal to the implanted fibers. As important...

...use a high fidelity polymerase to minimize mutations. The fidelity of the polymerase becomes more important...You should select an annealing temperature based on the melting temperature (Tm) of the portion of the primer... million bp depending on the polymerase used. No matter which polymerase you use, it is important that...restriction site to improve cutting efficiency. You can generally add any 6 bases, but you should ensure that the...with 18bp of homology to the ORF). We now need to generate the reverse-complement of this sequence so that...that we can successfully amplify the ORF. You can generate the reverse-complement using existing software...you can use a pretty wide range of annealing temperatures, but you may need to increase your primer length...

...localization, interaction, and expression. However, one caveat of FP-protein fusions (FP-chimeras) is that ...that they undergo normal protein turnover. FP-chimeras are continuously synthesized and degraded within the...488nm blue light. The T203H mutation in PA-GFP generates a chromophore population that is more neutral,...developed red fluorescent PA-FPs by performing several rounds of random mutagenesis screening for enhanced...formation of mEosFP’s chromophore requires a temperature below 30°C and this is not ideal for experiments...tissue than the UV light used by other PA-FPs. Several other irreversible PA-FP spectral variants have...choosing a PA-FP for your experiment, there are several parameters to consider, such as brightness, photostability...

... a DOI referencing primary literature, and every effort is made to moderate and cross-check all data with...affected by environmental conditions (such as temperature, pH, fusion protein, etc...).  There are many... Addgene blog – outlining various important considerations when choosing a FP, but much of the primary... making such a decision remains spread across literature in publications that introduce these tools. ..., and I collaborated with him on a web-based interactive visualization of FP attributes (which turned ...that allows users to see when a given page has unmoderated changes and to view previous versions.  Currently... with published literature as soon as possible after submission. Using FPbase The most obvious use case...

...your incubation temperature. Some proteins aggregate at 95 °C, a common lysis temperature, so consider a...background noise. Temperature and time You may need to optimize your antibody incubation temperature and time.... variants Check literature Posttranslational modifications Check literature Highly charged...said, normalizing to total protein expression is preferable, and more accurate, than normalizing to ubiquitously...recombinant multiclonal antibodies to increase overall signal from the primary antibody.  Pro tip! If ...common conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies... including trying longer incubations at both temperatures if needed. Washes Check your antibody manufacturer...

...coming of age. Molecular Therapy : the Journal of the American Society of Gene Therapy, 17(8), 1316–1332. https...can be an advantage for researchers looking to generate long-lasting changes in cells. However, for some...plasmids. For simplicity, we’ll use the second-generation system as an example.  The lentiviral transfer...integration, two types of circular episomes can be generated through either homologous recombination or non-homologous...integrase-deficient lentiviral vectors? There are several reasons why you might choose an IDLV over a lentiviral...clinical settings, as evidenced by retroviral gene therapy trials for X-linked severe combined immunodeficiency... 2000s. These trials unfortunately resulted in several patients developing T-cell acute lymphoblastic ...

...we’ll go through some of the most important considerations to make when planning and performing an IF ...IF experiment. What is immunofluorescence? Generally speaking, IF is an application in which antibodies...iBiology). IHC and ICC are essentially just more general terms that can also refer to assays using non-fluorescent... visualize their target. These antibodies are generated against a specific antigen and then conjugated... more common is that secondary antibodies are generally polyclonal, which means that multiple secondary...amplification. Another reason is cost effectiveness. Generating and validating primary antibodies is a time consuming...antibodies that exist and you can begin to see why generating fluorophore-conjugated versions for each primary...

...biotinylation, the discovery of the biotin-avidin interaction and the uses of biotinylation in molecular biology... and energy transduction. Biotinylation is the general process by which biotin is added to a protein or...protein (BCCP). Discovery of the biotin-avidin interaction   Figure 1: Biotin is enriched in egg ... of the strongest noncovalent protein-ligand interactions found in nature. Once the bond is formed it ...it is unaffected by changes in pH, temperature or organic solvents. Uses of biotin in molecular biology...scientists began to exploit biotin and its strong interaction with avidin for molecular biology and biotechnology...that are coated with avidin. The biotin-avidin interaction has also been adapted for use in immunohistochemistry...