When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.
When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human fibroblast-like synoviocytes
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human islets of langerhans
Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human dermal fibroblasts
Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - primary human cardiac fibroblasts
Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - immortalized Human blood cord
Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human CD14+ monocytes
Get tips on using pSilencer 2.1-U6 Hygro to perform shRNA gene silencing Human - Neuroblastoma cells (SH-SY5Y) Beclin 1
An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.
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