We narrowed to 791 results for: REP

... can then be purified for subsequent library preparation and high-throughput sequencing. Improvements ...lab Very recently, the Henikoff lab published a preprint paper outlining improvements to the initial CUT... rather than to rely on nuclei extraction. They report that this change offers a significant improvement... in terms of initial steps of cell or tissue preparation.   All in all, CUT&RUN offers a great alternative...

... the most frequently requested vectors in our repository and will discuss some of the features you may...more traditional approach of an autonomously replicating vector that carries either a wild-type version...Some of the more popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. ...animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g...

... perspective – a plasmid-containing cell must replicate the plasmid in addition to its own chromosomal...protein synthesis Bactericidal 7.5-50 ug/mL Streptomycin aminoglycoside Inhibits initiation of protein...working concentrations. For instructions on how to prepare antibiotic stocks, see Addgene's Reference Page...solution. Storing aliquots at -20oC and avoiding repeated freeze/thaw cycles will keep most antibiotics ...

...Cas9 is fused to a transcriptional activator or repressor to create a reversible effect. These approaches... several libraries are offered as lentiviral preparations to save you a few steps! To check whether a ...library you’re interested in is available as a viral prep, look under “Ordering” to see what options are offered...resources on the Addgene blog Viral Vectors 101: Preparing Pooled Libraries Pooled Library Amplifications...

...frequently denature the proteins as part of the sample prep. This involves using heat and/or chemicals to break...visualize an abstract protein during your sample prep process, imagining how it starts and if any steps...validated for an assay that uses similar sample preparation - for instance, if you cannot find an antibody...for recognizing the protein following your sample prep steps; ie, you may only need to validate on denatured...

...sequence information, leading to problems with reproducibility and an inability to improve upon these critical...provide renewable reagents with reliable and reproducible production. However, even clonal hybridoma cell...responsibility of the authors and does not necessarily represent the official views of the National Institutes ... References Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature...

...challenging and requires specialized equipment. Reporters for studying gene expression have similar drawbacks...added to the palette. RUBY is a bright red pigment reporter whose expresion is visible to the naked eye in... enzymes and a spontaneous step. (B) RUBY is a reporter consisting of these three enzymes expressed under...References  He, Y., Zhang, T., Sun, H. et al. A reporter for noninvasively monitoring gene expression and...

...rejoining supercoiled DNA ends to facilitate replication. The TOPO technique specifically uses Vaccinia...limit (lower transformation efficiencies have been reported with longer incubation); however, you may need...fairly quick, make sure you stay organized and prepare everything you need for the next step before proceeding... Perform Site Directed Mutagenesis by PCR Use REPLACR Mutatgenesis to Easily Generate Insertion and Deletions...

...some unexpected results. Order Ready to Use Viral Preps from Addgene Measuring the pH of our samples: What...closed dry ice storage container for 48 hours (to replicate how our viruses are shipped), where they would...., 2014). Based on our results and the results reported elsewhere, we recommend that once you receive ... CRISPR User) Tips for Titering Your Lentivirus Preps Additional Resources on Addgene.org Check out ...

...and Addgenie Angela Abitua hosted a virtual Reproducibility for Everyone workshop at TAGC with the three...engaged through a computer screen. For the Reproducibility for Everyone Workshop, Abitua and the other...knowing you're suppose to draw Spongebob? The Reproducibility for Everyone Workshop included this interactive... activity during their session. Image from Reproducibility for Everyone.  While this type of interaction...

...species of bacteria. The stain colored one type, Streptococcus pneumoniae, while the alcohol used in the staining...Klebsiella pneumoniae. After realizing this was a repeatable phenomenon, Gram began using it as a classification... positive bacteria include staphylococci and streptococci. Gram negative bacteria, or diderms, have a ...specific procedural instructions, including timing, preparing the smear, and mixing the stains and solutions...

...The transgene sequence is flanked by long terminal repeat (LTR) sequences, which facilitate integration of... be used interchangeably. Are γ-retroviruses replication incompetent? Yes, as with lentiviruses, the genes...Subcomponents: -U3 -R -U5 LTR; Long terminal repeats; U3-R-U5 regions found on either side of a retroviral...activation of viral genomic RNA transcription. R; Repeat region found within both the 5' and 3' LTRs of ...

...immunotherapy, as well as new approaches to reprogramming tumor-associated inflammation for therapeutic...reflect how cancer/immunology researchers are preparing and carrying out this multi-pronged attack. Inflammation...This immunosuppressive environment in turn may represent a favorable state for tumor progression (Ref 2...potential to become an open-source pipeline and data repository for cancer researchers. Go open-source! Impressions...

... et al used four oligos that consist of direct repeats and crRNA.  Similar to a jigsaw puzzle, the oligos... reverse the orientation of the crRNA’s direct repeats when expressing the Cpf1 multiplex system from ...virus.  Reversing the orientation of the direct repeat protects the (+) stranded lentivirus RNA from Cpf1...This undoes the original reversing of the direct repeats, leading to expression of a crRNA array that can...

... bioinformatic screens that looked for a CRISPR repeat array near a putative effector nuclease. The Cas13d...localization signal (NLS). In a HEK293 mCherry reporter assay, CasRx and AdmCas13d produced 92% and 87%...processed gRNA array (22 nt spacer with 30 nt direct repeat) mediating potent knockdown. Multiplexing from ...Addgene Blog: Learn about CRISPR RNA Editing with REPAIR Get tips for using Cas9 Transcriptional Activators...

...thing’ could be a neurotransmitter, such as norepinephrine, or an electrolyte such as calcium. This varied...transducer will generate a numerical value which represents the amount of analyte present in the system at...These sensors have been tested in vivo, in tissue preps, in multiple species, and in cell culture models... several years!). Biosensors tend to be more reproducible when used as stable systems, rather than transiently...

...spend more prep time on an experiment than it is to rerun an experiment, so consider your prep an investment... used to constantly underestimate time needed to prep for an experiment (labeling tubes never takes longer...harder to start earlier, but simply moving all my prep work to the day before, which got rid of the issue...

... are first injected with a small peptide that represents a short, unique amino acid sequence found in ...cells and antibody-producing B cells that cannot replicate, meaning that only hybridomas will survive this...monoclonal antibodies, they also lead to more reproducible experimental results. When you are originally...about the Developmental Studies Hybridoma Bank, a repository for hybridomas  Learn about monoclonal antibodies...

...the crRNA also contains a ~20 bp repeat region on its 3’ end. This repeat region associates with the trans-activating...is 3’ phosphorothioate linkage, where a sulfur replaces a nonbridging phosphate oxygen. Other useful backbone...Addgene's CRISPR resources Request ready-to-use viral preps of select CRISPR lentiviral plasmids Additional...

...sites (thrombin and TEV), and C-terminal tags (StrepII, 3xFLAG, 6xHIS, MBP, Myc). These constructs include...manually transforms, innoculates, and performs DNA minipreps for small batches of samples. As soon as that ...that process is complete, QC scientists prepare Sanger reactions by mixing this isolated plasmid DNA with... each group of plates will have select samples prepared and screened by NGS to spot check the samples,...