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Get tips on using Bacteria Live/Dead Staining Kit to perform Live / Dead assay bacteria - Corynebacterium glutamicum

Get tips on using LC3B Antibody Kit for Autophagy to perform Autophagy assay cell type - Corticospinal motor neurons

Get tips on using Gibco™Neurobasal™ Medium to perform 3D Cell Culture Media hiPSC-derived cortical organoids

Get tips on using Gibco™DMEM/F-12 to perform Stem cell Differentiation media hESCs differentiation into cortical neuroepithelium (NE)

Get tips on using Gibco™Glasgow's MEM (GMEM) to perform Stem cell Differentiation media hESCs differentiation into cortical neuroepithelium (NE)

Get tips on using Gibco™KnockOut™ Serum Replacement to perform Stem cell Differentiation media hESCs differentiation into cortical neuroepithelium (NE)

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 LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy to perform Live / Dead assay bacteria - Corynebacterium glutamicum

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

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.