We narrowed to 855 results for: NTS

... viral components or the transgene itself are used to quantify the number of integration events. Another...common assays for physical titer are direct p24 measurements via ELISA and qPCR for viral RNA. Measuring ...quantified using qPCR primers targeting specific viral components such as LTRs, gag, WPRE, antibiotic resistance-genes...genome and a cell that has had multiple integration events. In addition, promoter choice can also impact transgene... approach can discriminate multiple integration events, reducing the possibility of underestimating the...titering as this will better represent future experiments. The aforementioned titering options all have...of infection (MOI) for predicting gene transfer events." Genetic vaccines and therapy 2.1 (2004): 1. PubMed...

...contamination free environment for successful experiments: To clean air and beyond Turn on the hood for...wipes) before and after your work. Keep your contaminants away While it’s likely that you’re the main ...main source of contamination for your experiments, the bright side is that you also have the power to take...aerosols that may enter the hood. Planning your experiments thoroughly before entering the hood is another...microscope will help you identify bacterial and yeast contaminants. Set up a routine to split cells Don’t be lazy...rendering their continued use inadvisable for experiments requiring reproducible results. Most cell culture...should not be used for important transfection experiments right after they’ve been thawed. Additional passages...

..., safety requirements and expression limitations may be too constraining for some experiments, particularly...that often affect which is chosen for certain experiments. AAV can be safely handled at BSL-1, while lentivirus...vectors is how the cargo — in this case, the CRISPR components — are expressed. Unlike lentivirus, cargo delivered...AAV genome can integrate at double-stranded break points caused by CRISPR-Cas9 editing. The good news is...the amount of cloning, another option is to use variants of Cas9, like split Cas9, or a smaller Cas9 ortholog...tissue-specific diseases has grown exponentially, aided by advancements in in vivo CRISPR screens. AAV vectors have ...have played a large role in these advancements, due to their many benefits for in vivo models. Over the past...

...introduce their students to this otherwise invisible molecular world. For the students, the models force...hand-deliver the model, the following sequence of events plays itself out: The researcher breaks eye-contact...membrane of the endoplasmic reticulum. And graduate students, momentarily forgetting that the physical model...Subscribe to CRISPR Blog Posts How do educators and students use molecular models?   We created the Center ...instances of focused attention become magical teaching moments. The models function as thinking tools – eliciting... Team modeling program in which small teams of students work closely with local researchers to learn about... designed to make the molecular world real for students. Many thanks to our guest blogger Tim Herman!...

...generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters et al., 2019 with permission...using the bacterial integrative and conjugative elements in Bacillus subtilis (ICEBs1) system. ICEBs1 carries... Top: A Tn7 transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes...lacking DAP select against donor bacteria and for recipients with an integrated copy of CRISPRi. Bottom: An... An ICE element containing the CRISPRi components is transferred to recipient bacteria by bi-parental ...suited for genome-wide CRISPRi library screening experiments where maintaining library diversity is important...rates are only suited for single-gene knockdown experiments. Mobile-CRISPRi applications Mobile-CRISPRi expands...

... mutually exclusive elements. Labs have successfully utilized 2A and IRES elements within the same multicistronic...for research purposes are described below. IRES Elements Translation in eukaryotes usually begins at the...translation initiation independent of the 5’ cap.  IRES elements are very useful and commonly found in bicistronic...however, they do have some disadvantages. These elements are quite large (500-600 bp) and may take up precious...express more than two genes at a time using IRES elements. Further, scientists have reported lower expression...features and spend more time developing the clever experiments and innovative solutions necessary for advancing...ITAMs ensures effective negative selection and prevents autoimmunity. Holst et al (Nat Immunol. 2008 Jun...

...colors and 10 points to keep in mind when choosing an FP (or two) for your upcoming experiments. Fluorescence...FPs just keeps getting larger with hundreds of variants available. Read on to familiarize yourself with...emission wavelenght range).  By mutating GFP, the variants blue FP (BFP), cyan FP (CFP), and yellow FP (YFP...YFP) were derived. For a breakdown of GFP, it's variants, and their relevant mutations, check out Marcy's...Bimolecular Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing a fluorescent protein...you are planning to express the FP in acidic environments (e.g. yeast cytosol, which is slightly acidic...with very comprehensive data sheets on many FP variants Plasmids 101: Green Fluorescent Protein (GFP) ...

...sample is pure and free of any contaminants that could affect the measurements. You'll need to prepare the... or very similar buffers if not, since buffer components can affect the final readout. When preparing ...duplicate or triplicate. The closeness of the data points generated by replicate values provides useful information...diluted standard is plotted against its OD560 measurements and the trendline y = 0.6561x + 0.019 calculated...predicate values and measures how well the data points fit the trendline. R2 ranges from 0 to 1 with 1...approaches the minimum and maximum thresholds. If the measurements for the unknown sample fall outside of the dynamic... to set yourself up for success!  Study the instruments and specific assays you will be using ahead of...

...in high pH environments and is less susceptible to degradation in agar plates. This prevents satellite ...less stable at high temperatures and in acidic environments.   Carbenicillin   Carbenicillin, colloquially...genomes. If you need a plasmid that can go between plants and bacteria, it’s a great option! Otherwise, since...eukaryotic cell culture, yeast, bacteria, and even plants! The zeocin resistance gene is present in bacteria...et al). Thus, it may not be a good choice for experiments in which off-target mutations are undesirable..."Check icon" %} Zeocin Excellent No Bacteria, plants, and eukaryotes Not Tn5-containing cells {% icon...temperature and pH on the stability of antimicrobial agents in MIC trays. Journal of Clinical Microbiology,...

...(genomic DNA and RNA), proteins and chemical contaminants that have been used during the purification ... indicators of DNA purity are the absorbance measurements at 230, 260, and  280 nm. The absorbance ratios...scientists an indication of purity versus protein contaminants. The 260 and 230 nm ratios should fall ~2 - ...provide an indication of purity versus chaotropic agents as guanidine thiocyanate and guanidine hydrochloride...extraction of enough plasmid to last for multiple experiments. Another fundamental aspect for transfection ... being free of any nucleic acid and chemical contaminants, these DNA samples must also be entirely endotoxin...ultimately affects the quality and outcomes of experiments. For these reasons commercially available kits...