DNA isolation / purification Cells Immortalized cell lines

Get tips on using Live/Dead cell Staining Kit II to perform Live / Dead assay mammalian cells - NIH/3T3

Get tips on using Live-Dead Cell Staining Kit (BioVision) to perform Live / Dead assay mammalian cells - NIH/3T3

Get tips on using Live and Dead Cell Assay (Abcam) to perform Live / Dead assay mammalian cells - mouse keratinocytes

Get tips on using CometChip Electrophoresis Starter Kit to perform DNA Damage Assay Human bronchial epithelial cells (hBE)

Get tips on using Comet SCGE assay kit to perform DNA Damage Assay Human bronchial epithelial cells (hBE)

Get tips on using PrimaPure Porcine Endothelial Cells Growth Medium to perform Mammalian cell culture media PAOEC

Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

Get tips on using Qubit dsDNA HS Assay Kit to perform DNA quantification MCF10A breast epithelial cells

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.