...We often think of DNA as inert. It generally stays put, which makes it easy to locate in a genome. But...mobile DNA, called a transposon, that’s a bit hyperactive and likes to jump around from one location in... at rates similar to retroviral vectors. The hyperactive version of the SB transposase, SB100X, is ~100...100-fold more efficient compared to first generation SB transposase (Mátés et al., 2009). hySB100x improves...human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold more active...transposon insertion sites are detected with next generation sequencing, and analyzed to identify which insertions...Transgenic animals Transgenic animals are often generated by directly injecting DNA into the pronuclei of...

...Newman-Keuls method – More powerful. Moderate false negatives, moderate false positives. Duncan’s multiple...is being experimentally manipulated. Below are several core factors of statistics relevant for many downstream...Parametric tests  Statistical tests fall into two general categories: parametric and non-parametric. In this...variable. A two-way ANOVA could test the effect of temperature and humidity on an outcome such as total crop... one-way ANOVA can test either the effect of temperature or humidity on crop production.   MANOVA The ...example, a one-way MANOVA could test the effect of temperature on total crop production and the production window...A two-way MANOVA could measure the effect of temperature and humidity on total crop production and the...

...translation by generating a frameshift mutation. The majority of cellular repair events will generate these types... a less frequent DNA repair mechanism in general. On average, it accounts for less than 10% of DNA repair...family member which generates sticky ends upon cutting. Cas12a may promote HDR by generating ssDNA overhangs...Taking the road less traveled and generating a knock-in cell line instead of a knock-out? We’ve got you...editing Targeted genome editing events fall into two general classes: knock-in and knock-out. The goal of a ...exogenous DNA can be used as well. Below are some considerations for designing donor DNA for successful HDR ...viability of your cells. Enrich S-phase cells HR operates predominantly in S and G2 phase of the cell cycle...

... the notation works? The + operator denotes intersection and the – operator denotes set difference. That... ChiRP-seq RNA Hi-C DNA (genome-to-genome interactions) DNase-seq DNA (regions that are accessible... the foundation of modern drug discovery and therapeutics. We’ll have more to say about clinical applications...Poisson statistics, each sub-population has, on average, one distinct enhancer disrupted. As our ESCs die...differentiate over time, we periodically use next-generation sequencing to measure the relative proportion...apply this knowledge to create targeted disease therapies with minimal side-effects on healthy cells and...Hypersensitive Sites and Intergenic CRISPRs Yours truly generated the DHS data set by starting with all of the DHS...

...including tips, advantages, and disadvantages of this overall rewarding role. Learn about Other Addgenies and...role in technical support? It takes some time (several years) to build up comprehensive lab experience...You will also help troubleshoot products, give general experimental guidance, and resolve complaints about...willingness to help others with their problems, a general positive attitude, and an open-minded personality...any incoming requests or problems - there is considerable diversity in the types of questions customers...will do well in a technical support role. What general characteristics do jobs in technical support have...covered by the lab based team as part of their general job duties. Bigger companies have dedicated technical...

... 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...