siRNA / RNAi /miRNA transfection Human Cells

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Get tips on using FlexiTube GeneSolution GS7052 for TGM2 to perform siRNA / miRNA gene silencing Human - Caki-2 TGM2

Products Qiagen FlexiTube GeneSolution GS7052 for TGM2

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.

Cellular assays Live / Dead assay mammalian cells MDA-MB-231 human breast cancer cells

Get tips on using Orai1 Rat siRNA Oligo Duplex (Locus ID 304496) to perform siRNA / miRNA gene silencing Rat - RBL-2H3 Orai1

Products OriGene Orai1 Rat siRNA Oligo Duplex (Locus ID 304496)

Get tips on using ON-TARGET plus Mouse Becn1 (56208) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - 4T1 BECN-1

Products Horizon Discovery Ltd. ON-TARGET plus Mouse Becn1 (56208) siRNA - SMARTpool

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Proteins ChIP Human Kupffer Cells

Get tips on using ON-TARGETplus Rat Rac1 (363875) siRNA - Set of 4 to perform siRNA / miRNA gene silencing Rat - MTLn3 Rac1

Products Horizon Discovery Ltd. ON-TARGETplus Rat Rac1 (363875) siRNA - Set of 4

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.

RNA RNA isolation / purification Cells primary human chondrocytes

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.

RNA RNA isolation / purification Cells primary human osteoblasts

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.

RNA RNA isolation / purification Cells primary human preadipocytes

Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,

DNA DNA isolation / purification Cells Primary cells Human primary keratinocytes

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