siRNA / miRNA gene silencing Human MDA-MB-453

Get tips on using EZ4U - Cell Proliferation Assay to perform Live / Dead assay mammalian cells - GH3

Get tips on using Cellstain-Double Staining Kit to perform Live / Dead assay mammalian cells - A549

Get tips on using Cellstain-Double Staining Kit to perform Live / Dead assay mammalian cells - K562

Get tips on using Viromer® RED to perform DNA transfection Mammalian cells - Immortalized cell lines HEK293

Get tips on using Viromer® RED to perform DNA transfection Mammalian cells - Immortalized cell lines HeLa

Get tips on using PolyFect Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines CHO

Get tips on using Viromer® RED to perform DNA transfection Mammalian cells - Primary cells Rat astrocytes

Get tips on using PolyFect Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines HEK293

RNA quantification for appropriate concentration and quality (260/280 ratio) is an important step before downstream analysis (including sequencing, RT-qPCR, etc.). Having insufficient RNA quantities or a high salt or phenol in the RNA product can lead to variable or irreproducible downstream results. The various methods used for RNA quantification include: 1. UV spectrophotometric (challenges include: low sensitivity, cannot distinguish between nucleic acid species), 2. Fluorescence-based (challenges include: requires standards, cannot measure amplifiability, not sequence-specific), and 3. RT-PCR (challenges include: requires standards, time-intensive, costly). In order to overcome these challenges, and also to ensure the proper quantification and quality control for RNA product, it is important to use at least two or more methods in order to discard any inconsistencies. Using standards for calibrations increases the sensitivity range for RNA detention (fluorescence- and RT-PCR-based methods). When using RT- PCR, it is important to choose correct primers, aligning to the desired site on the template and of appropriate product length, along with positive, negative and loading controls. It is also important to have at least two primer pairs in order to confirm results.

RNA quantification for appropriate concentration and quality (260/280 ratio) is an important step before downstream analysis (including sequencing, RT-qPCR, etc.). Having insufficient RNA quantities or a high salt or phenol in the RNA product can lead to variable or irreproducible downstream results. The various methods used for RNA quantification include: 1. UV spectrophotometric (challenges include: low sensitivity, cannot distinguish between nucleic acid species), 2. Fluorescence-based (challenges include: requires standards, cannot measure amplifiability, not sequence-specific), and 3. RT-PCR (challenges include: requires standards, time-intensive, costly). In order to overcome these challenges, and also to ensure the proper quantification and quality control for RNA product, it is important to use at least two or more methods in order to discard any inconsistencies. Using standards for calibrations increases the sensitivity range for RNA detention (fluorescence- and RT-PCR-based methods). When using RT- PCR, it is important to choose correct primers, aligning to the desired site on the template and of appropriate product length, along with positive, negative and loading controls. It is also important to have at least two primer pairs in order to confirm results.