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 Anti-EpCAM antibody (ab71916) to perform Immunohistochemistry Mouse - EpCAM
Get tips on using GDNF RECEPTOR ALPHA 1 to perform Immunohistochemistry Mouse - GFRA1
Get tips on using CD31/PECAM-1 Antibody to perform Immunohistochemistry Mouse - CD31
Get tips on using Anti-CD31 antibody (ab28364) to perform Immunohistochemistry Mouse - CD31
Get tips on using Anti-Choline Acetyltransferase Antibody to perform Immunohistochemistry Mouse - ChAT
Get tips on using Anti-βIII Tubulin mAb to perform Immunohistochemistry Mouse - TUBB3
Get tips on using Goat antibody to Calretinin to perform Immunohistochemistry Mouse - Calb2
Get tips on using Anti-Myeloperoxidase antibody (ab9535) to perform Immunohistochemistry Mouse - MPO
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment