protein-expression-and-purification-bacteria-escherichia-coli-p19

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Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

RNA siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line) HSPA5 (GRP78)

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

RNA siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line) SIRT1

Get tips on using FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme to perform TUNEL assay cell type - 3T3 L1 mouse adipose tissue

Products Millipore FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme

There are a plethora of detection methods of cell cytotoxicity and proliferation by 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 Cell cytotoxicity / Proliferation assay cell type MCF-7

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.

DNA DNA methylation profiling Whole genome profiling HCT116, HTC15 human colon cancer cells

Get tips on using QIAGEN Protease (30 AU) to perform Digestion during DNA preparation

Products Qiagen QIAGEN Protease (30 AU)

DNA DNA quantification Human Colorectal aenocarenoma (SW48) - paraffin embeded

Cell culture media 3D Cell Culture Media Murine colonic organoids

Cellular assays Cell line authentication Colon cancer cell line LoVo

Cellular assays Cell line authentication Colon cancer cell line RKO

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