protein-expression-and-purification-mammalian-cells-hek-293-mt-pa

miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.

Get tips on using mRFP-green fluorescent protein (GFP)-LC3 adenoviral vector to perform Autophagy assay cell type - Macrophages

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 Aurum™ Total RNA Fatty and Fibrous Tissue Kit to perform RNA isolation / purification Tissue - Human Uterus

Get tips on using Blu10 Plus (BLUltra) Prestained Protein Ladder（6.5 to 270 kDa） to perform Protein Ladder Prestained

Get tips on using Blue Prestained Protein Marker, Broad Range (11-250 kDa) #59329 to perform Protein Ladder Prestained

Get tips on using Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028) to perform Protein Ladder Prestained

Get tips on using jetPEI®-HUVEC to perform DNA transfection Mammalian cells - Primary cells HUVEC

Get tips on using NucleoSpin® Gel and PCR Clean-up to perform DNA gel extraction / PCR product purification Product size > 15Kb

Get tips on using MasterPureTM-Yeast DNA Purification kit to perform DNA isolation / purification Yeast - Pichia pastoris