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

Get tips on using 8 µm Chemotaxis Assays, 96-Well Format to perform Cell migration / Invasion cell type - MCF-10A

Get tips on using 8 µm Chemotaxis Assays, 24-Well Format to perform Cell migration / Invasion cell type - PANC-1

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Cell cytotoxicity / Proliferation assay cell type - FADU

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Cell cytotoxicity / Proliferation assay cell type - HeLa

Plasmid isolation is an important technique in molecular biology or any kind of genetic editing. It involves amplifying plasmids overnight by transforming them into competent bacterial cells. The desired colonies of these bacteria can then be grown in shaker cultures, at appropriate shaking speed, oxygen availability and temperature. These liquid cultures can then be ultracentrifuged to pellet the bacteria, which are then used for plasmid isolation. The bacteria are first resuspended in a buffer, then lysed, neutralized, purified in a column, eluted, precipitated with ethanol and then resuspended. During plasmid isolation, it is important to lyse cells quickly because lysing bacteria for too long may lead to irreversible denaturing of the plasmid. Usually, alkaline lysis is used for isolation because it is a mild treatment. It isolates plasmid DNA and other cell components such as proteins by breaking cells apart with an alkaline solution. Precipitation removes the proteins, and the plasmid DNA recovers with alcohol precipitation. Resuspension and lysis buffers should be mixed thoroughly in order to prevent the DNA from breaking into smaller fragments. This is because broken gDNA can reanneal and remain in the solution, without binding to the column.

RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.

Get tips on using Gibco™ DMEM, high glucose to perform Mammalian cell culture media PCAEC

DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Cell cytotoxicity / Proliferation assay cell type - THP-1

Get tips on using LSI VetMAX™ Bluetongue Virus Real-Time PCR Kit, BTV1 typing - IAH to perform Cell Culture Contamination Detection Kit Virus