Access Array System for the Ion Torrent PGM Sequencing System: User Guide
Chapter 4: Sample Quantitation and Normalization
Sample Quantitation
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Sample Quantitation
Quantitate the samples by fluorimetry, using the Quant-iT PicoGreen dsDNA Assay Kit.
Follow the manufacturer’s instructions.
Sample Normalization
The following recommendations apply to human genomic DNA samples. For other samples,
the recommendations on sample concentrations may be different. Contact Technical
Support for assistance in this case.
• If the sample concentration is 50 ng/L gDNA input (a total of 50 ng gDNA), the sample is
ready for amplification on the LP 48.48 IFC. For germline mutation, you might be able to
use as little as 525ng/L.
• If the sample concentration is below 50 ng/L gDNA, we recommend concentrating the
sample before amplification takes place on the Access Array.
• If the sample concentration is above 50 ng/L, we recommend diluting the sample to
50 ng/L using DNA Suspension Buffer before proceeding.
Use the following formula to determine the correct volume of DNA Suspension Buffer
required to dilute each sample to 50 ng/L:
Y = X (B/50 – 1)
Where X is the volume of the original sample (L) to be used in the dilution
Y is the DNA Suspension Buffer volume (L) needed to dilute X L of the original sample
to 50 ng/L
B is the sample concentration (ng/L) measured by fluorimetry
50 is the desired sample concentration (ng/L)
For example: If a 10 L sample (X = 10 L) has a concentration of 200 ng/L
(B = 200 ng/L):
Y = 10 L * ((200 ng/L)/(50 ng/L) –1)
Y = 30 L
Therefore: Dilute 10 L of the 200 ng/L sample in 30 L of DNA Suspension Buffer to
obtain a 50 ng/L sample concentration.
• Normalize all sample concentrations to 50 ng/L.
Samples are now ready for amplification using the LP 48.48 IFC.
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