RNA isolation / purification Plants

Get tips on using RPM Yeast Plasmid Isolation Kit, 100 preps to perform Plasmid Isolation S. cerevisiae

Get tips on using Pan Monocyte Isolation Kit, human to perform Cell Isolation Monocyte

Get tips on using Classical Monocyte Isolation Kit, human to perform Cell Isolation Monocyte

Get tips on using MACSprep™ PBMC Isolation Kit, human to perform Cell Isolation PBMC Isolation

Get tips on using QIAamp MinElute ccfDNA Mini Kit (50) to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using QIAamp DSP DNA Blood Mini Kit to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using EZ1 DSP DNA Blood Kit (48) to perform DNA isolation / purification Tissue - blood / plasma

Get tips on using QIAamp DNA Blood Mini QIAcube Kit to perform DNA isolation / purification Tissue - blood / plasma

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.