We narrowed to 855 results for: NTS

...technology itself is constantly improving. Cas9 variants have been modified for genome editing, activating...specificity of the Cas9 nuclease with engineered variants: eSpCas9, SpCas9-HF1, & HypaCas9. The off-target...substitutions throughout the groove in 32 separate Cas9 mutants. When tested for their ability to cut the EMX1 ...embryonic kidney (HEK) cells, 11 of these initial mutants retained the on-target activity of wt Cas9 while... of these mutations and tested the combination mutants for their on-target and off-target activity at ...locations in HEK cells. Further testing of two of the mutants, SpCas9(K855A) and eSpCas9(1.1), followed by a ...double strand breaks (BLESS), revealed that these mutants do not cause off-target effects at unanticipated...

... compatible with detergents like SDS, while BCA assays are not with denaturing agents like DTT or β-mercaptoethanol...other hand, they are not as sensitive to lower amounts of protein, and can require more antibody or loading... loading buffer will not contain any denaturing agents. However, you’ll still have to lyse your proteins...protein. Proteins located in membrane-bound subcompartments, like the mitochondria or nucleus, will likely...gradient gel, the decision is probably already made: gradients are extremely difficult to handcast, so unless...blots. If you need to preserve sample, check the ingredients before using, as some contain Tween-20, which...reaction rate and are compatible with antimicrobial agents. Chemiluminescence kits When activating your enzyme...

...may undergo genomic rearrangements during replication. To avoid genomic rearrangements, try amplifying the...contact A variety of reagents can increase transduction efficiency. These reagents typically enhance the...working with virus: 1. Viral vectors undergo DNA rearrangements. Depending on the composition of the viral...viral vector with bacteria designed to minimize rearrangements such as NEB Stable or use a standard strain...ensuring viral production is to perform PCR against components of the viral vector using the harvested viral... you time, and allow you to standardize your experiments later. Stay tuned to the Addgene blog and be ... pages to get up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A...

...only will these mStayGold variants be valuable tools for your next experiments, they will also likely inspire...genome editing for primary cells Monomeric StayGold variants AAV.CAP-Mac for systemic gene transfer in primate... Breaking up is hard to do: Monomeric StayGold variants By Mike Lacy In 2022, Atsushi Miyawaki and colleagues...stability. Here’s a quick overview of these new variants:   Figure 2: Inner mitochondrial membranes...The Miyawaki lab generated a series of StayGold mutants and identified a triple mutant, Y187F, R144I and...Addgene here!   Ando, R., et al. (2023). StayGold variants for molecular fusion and membrane-targeting applications...gene delivery capsids previously identified in rodents have not been widely successful in primates. The...

...structures or other local genomic elements (Hazan and Bester 2021). Other Cas variants Another option is CRISPR...ncRNA (Jiang et al. 2014).  Simple knock-out experiments are trickier when it comes to long non-coding... et al. 2018). However, removing large lncRNA fragments with multiple gRNAs poses the risk of disrupting...can be minimized by disrupting lncRNA regulatory elements that are crucial for transcription and downstream...or introduced, into the break sites. Knock-in experiments are important for visualizing ncRNA localization...long ncRNAs is impractical. Instead, tags or stop elements can be inserted to disrupt the transcription of...CRISPR/Cas9 system is not effective for your experiments, there are plenty of other CRISPR-based options...

...to extract without harsh reagents and patience. Plant expression systems Plants provide a cheap and low-tech...through the use of protein tags and separated from contaminants using a variety of purification methods. Deciding...systems can be scaled up for production of large amounts of protein. Some drawbacks of insect cell expression... systems. Bacterial expression systems When one wants to produce vast quantities of protein rapidly and... DNA vectors that can be used to produce large amounts of protein in bacterial cells: the pET, pRSET, ...recombinant proteins. Many cells from various types of plants such as maize, tobacco, rice, sugarcane, and even...share many of the same features and processing requirements as mammalian cell expression systems, including...

...Contamination by humans accounts for the largest source among those mentioned above. The contaminants can spread through...antibiotics and are broad-spectrum antibiotic agents. These agents can be used to wash the cell cultures. The...to the liquid or semi-solid medium (containing nutrients essential for mycoplasma growth). Infected cell...synthesized by eukaryotic cells. First, the kit components rupture the mycoplasma and then specific enzymes...for the lab to avoid bringing in environmental contaminants.  Figure 2: An Addgenie works in the...Photo by Kate Harten DeMaio. Cell lines and reagents Examine your cell line stock for mycoplasma contamination... subculturing. Avoid using expired products or reagents that have been stored for a longer time. Use fresh...

...repetitive genetic elements that are found in both prokaryotic and eukaryotic genomes.  Components of a transposon...around in the lab.  Let’s take a look at the key components of transposons, how they are classified, and ...target sites. Integration of most transposable elements (TEs) results in the duplication of this target...insertion.  Types of transposons Transposable elements are split into two classes based on their mechanism...the rest of this article. Class I Transposable elements (TEs): Retrotransposons  Class I TEs are also ...screens Transposons, by their nature, are mutagenic elements, which makes them a great tool for mutagenesis...developed the INTEGRATE (Insertion of Transposable Elements by Guide RNA-Assisted Targeting) system. INTEGRATE...

...DNA you’re purifying (plasmid, genomic, or DNA fragments from agarose).    In this article, we will go ...or agarose gel slices Product: plasmids or DNA fragments The plasmid purification and DNA gel extraction...Starting material: agarose gel slices Product: DNA fragments You might be wondering, “How is milk going to ...or even paper towels to purify DNA and RNA from plants, microbes, blood or mammalian cells in 30 seconds...into this mixture to absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids...placing the dipstick into a wash solution removes contaminants while nucleic acids stick around. Finally, purified...gel slices Product: purified PCR product or DNA fragments Maybe you’re not ready to give up DNA purification...

...permeabilization with detergents such as triton-x, saponin, or tween. Take care when using strong detergents like triton-x... and electronics systems, FACS instruments have specialized components that can divert a subset of cells...overcome these challenges and make these types of experiments easier and faster to perform. Introduction to...channel number based on intensity. Higher intensity events are assigned to higher channels. In a typical assay...assay a user will compare the shifting intensity of events. Since intensity positively correlates with expression... flow cytometry particularly well to complex experiments looking at a variety of different targets. If...for setting up antibody based flow cytometry experiments.       References and resources References:...