RNA / DNA Assesment
Starting material of the highest practical quality is important to ensure the best data is generated. Nanodrop (UV/Vis) measurements are good for assessing sample purity and estimating concentration, however, analysis by microfluidic devices (eg. Agilent Tapestation) are ideal for assessing RNA and DNA size and quality due to their high resolution and low sample usage.
All samples and libraries are required to be assessed prior to sequencing; the facility is able to perform this assessment at a reasonable cost for you if you don’t have access to these instruments. Where input concentrations are critical (or low by Nanodrop) a fluorescence measurement should be performed with an appropriate dye and reader (eg. Qubit) for accuracy. The table below shows the input requirements for some of the more commonly used Agilent sample assays.
AGILENT MICROFLUIDIC ASSAYS
|RNA/DNA Assessment||Sample Type||No. Samples / run||Input (ng/uL)||Supply (uL)|
|Agilent Tapestation High Sensitivity D1000 Screentape||DNA (50 - 1000 bp)||2 - 16||0.005 - 0.5||3|
|Agilent Tapestation RNA Screentape||Total RNA||2 - 16||50 - 500||3|
|Agilent Bioanalyzer RNA Picochip||Total RNA||up to 11||0.1 - 3||3|
|Agilent Tapestation Genomic DNA||DNA (200 - 60000 bp)||2 - 16||5 - 300||3|