Protein expression and purification Bacteria Escherichia coli

Get tips on using MEDOX-Easy Total RNA Extraction Reagent to perform RNA isolation / purification Tissue - Human Mouth

Get tips on using MagNA Pure Compact RNA Isolation Kit to perform RNA isolation / purification Tissue - Human Bronchi

Get tips on using EZ-RNA Total RNA Isolation Kit to perform RNA isolation / purification Tissue - Human Brain

Get tips on using SV 96 Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized T84

Get tips on using illustra™ RNAspin Mini Isolation Kit to perform RNA isolation / purification Cells - immortalized DH82

Get tips on using illustra™ RNAspin Mini Isolation Kit to perform RNA isolation / purification Cells - immortalized Vero

Get tips on using MagAttract 96 DNA Plant Core Kit (24) to perform DNA isolation / purification Plants - Leaves

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.

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human pulmonary arterial smooth muscle cells