The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.
Get tips on using Mouse Osteopontin/OPN Antibody to perform Immunohistochemistry Mouse - Spp1/OPN
TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.
Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Tissue - mouse liver
Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Tissue - mouse kidney
Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Tissue - mouse brain
Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Tissue - mouse thymus
Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Tissue - mouse spleen
Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Tissue - mouse aorta
DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.
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