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...may be thinking, well, these are mostly fairly standard molecular and cell biology techniques. What’s ...step can affect the optimization of another. For instance, a sample may need more or less cross-linking ...

... into the bacterial genome. REPLACR-mutagenesis starts out in familiar territory, but quickly diverges...recombination then joins the linear PCR products into stable circular plasmids - no nick repair is needed. Since...

...only 44% and 66% indel frequency with the use of standard single gRNA constructs. When PTG constructs with...<200 bp tRNA-gRNA takes up half the space of a standard pol III-gRNA, which will facilitate CRISPR/Cas9...

...intervening DNA (“pop-out” deletions) if NHEJ joins the distal ends together. Similarly, it may be possible to...References Bennardo N, Gunn A, Cheng A, Hasty P, Stark JM (2009) Limiting the Persistence of a Chromosome...

...several considerations. First, it is important to establish the type of audience you want to reach with your...and help you correct any language- and spelling mistakes. A manuscript that is well written looks more ...

...To assess TRIM21 toxicity, Clift et al. created stable cell lines overexpressing TRIM21, finding that ...which is necessary for effective STED), and be photostable over a broad pH range. The four biometric probes...

...However, transcription of thousands of genes is constantly occurring throughout a single cell, so these ... these structures can in turn lead to genome instability. Thus, it’s important to strike a balance between...

...DNA-protein interactions or to analyze chromatin states (CHIP-seq, FAIRE-seq, Cut & Run) and to detect ...interactions. To address this need, the Khavari lab at Stanford created the RNA-protein interaction detection ...

...resulting in GFP signal. With the addition of standards, the assay is semi-quantitative and can approximate...plasmids and suitability of various antibiotic resistance markers, promoters, and inducible gene expression...

...possible antibody conjugates (especially if you start to consider clinical applications). But there are...make the process quick and easy. Before getting started, be sure to check that your antibody’s buffer is...

.... The last step involves a chemical reaction to stain the cells that contain this mRNA-riboprobe-antibody.../79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver, C.C. Mello. Potent and specific genetic...

...Create promoters with sequences upstream of the start codon of retinal cell-type specific genes from mice...synthetic promoters. Legend: TSS: transcription start site. TFBS: transcription factor binding site. Image...

...vivo studies should always take DNA’s chromatin status into account (Fig 1a). The laboratory of Karmella...dCas9 expression and contain different antibiotic resistance markers. New CRISPR pooled library from Keith...

...that treat disease at the molecular level. For instance, if a disease is caused by a mutation in a single...developing AAV-based therapies. Furthermore, deeper understanding of disease pathogenesis will enable robust therapeutic...

...use electroporation, a technique that briefly destabilizes the cell’s outer membrane and allows the components...structures and processes and can lead to the better understanding of cell biology. We hope that these high-quality...

...considered "Cre-On" meaning that your gene of interest starts in the inverse/antisense "off" position and is ...employed for "Cre-off" (i.e. your gene of interest starts in the sense orientation and Cre is used to flip...

...et al., 2024) have highlighted how chromatin substantially affects prime editing outcomes across many genomic...with PRIDICT/PRIDICT2.0, ePRIDICT can help you understand how chromatin might influence editing results...

...light up endogenous genes of interest to better understand normal protein function. However, CRISPR protein...introduced directly into a Drosophila S2 cell line stably expressing Cas9. This PCR toolkit offers C- and...

...constructs are expressed transiently by transfection or stably impacts the success of BiFC experiments. In either...negative controls because their expression level, stability, and subcellular localization may differ from ...

...is carefully designed primers. The majority of mistakes are due to incorrect primer sequences! Primers...transduction, and uses electron cryo-microscopy to understand receptor function.     Follow them on Twitter...