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

Though DNA quantification is but one small step in the multifaceted DNA sample preparation workflow, it can have large implications on the performance and validity of conclusions drawn from downstream assays. Major challenges include accuracy, precision, reproducibility, and detection of present contamination. Among UV spectrophotometry, fluorescence and real-time PCR based methods, the quantification method should be chosen based on the requirement of the downstream assay.

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 ScriptSeq Complete Kit (Human/Mouse/Rat) to perform RNA sequencing Mouse - Bone marrow-derived macrophages (BMDMs)

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

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 Monoclonal Mouse Anti-Human Cytokeratin (Concentrate) Clone AE1/AE3 to perform Immunohistochemistry Mouse - Cytokeratin

Get tips on using Purified Mouse Anti-Human MLH1 Clone G168-728 (RUO) to perform Immunohistochemistry Human - MLH1

Get tips on using Human Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit to perform ELISA Human - ShhN

Get tips on using Human Retinol binding protein 4 ELISA Kit (RBP4) (ab108897) to perform ELISA Human - RBP4