Agilent 2019 - 18

Automated Nucleic Acid Sample Quality Control in NGS Workflows

Analytical Specifications
Table 1 summarizes the analytical specifications
of the D1000 and HS D1000 ScreenTape assays
for the 4200 TapeStation system and the
specifications of the DNA 1000 and HS DNA
assays for the 2100 Bioanalyzer system. The
analytical specifications of the ScreenTape
assays were previously systematically validated for
sensitivity, sizing, quantification, and molarity.
Results and Discussion
Molarity

Sample molarity is often used to determine the
load volume of NGS libraries for pooling before
sequencing. The calculation of molarity is based on
average size and concentration of library samples.
Thus, for accurate molarity results and successful
library sequencing, sizing and quantification must
be accurate. Excellent assay performance for NGS
library samples was shown previously for the
D1000 and HS D1000 ScreenTape assays. It was
also demonstrated that the assays are suitable
for sample quality control in a high throughput
sequencing environment.
The lower and upper size limits of the DNA assays
(Table 1) apply for maximum peak sizes of DNA
fragments. DNA libraries must match the assay
range with the whole sample size to fit between the

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Table 1. Analytical specifications of the D1000 and HS D1000 ScreenTape assays (4200 TapeStation system) and the DNA 1000 and HS DNA assays
(2100 Bioanalyzer system).

Agilent 4200 TapeStation System

Agilent 2100 Bioanalyzer System

D1000
ScreenTape Assay

High Sensitivity D1000
ScreenTape Assay

DNA 1000
Assay

High Sensitivity
DNA Assay

35-1,000 bp

25-1,000 bp

50-7,000 bp

Sizing accuracy*

±10 %**

±10 %

Sizing precision*

5 % CV

Quantitative range
(DNA fragments)

0.1-50 ng/µL

10-1,000 pg/µL

0.5-50 ng/µL*

5-500 pg/µL*

Quantitative range
(DNA smears)

5-100 ng/µL

0.5-15 ng/µL

0.1-10 ng/µL

±20 %***

±20 %*

25-500 bp: 15 % CV*

50-2,000 bp: ± 15 %*

500-1,000 bp: 5 % CV*

2,000-7,000 bp: ± 10 %*

Analytical specifications
Sizing range

Quantitative accuracy
Quantitative precision

0.1-1 ng/µL: 15 % CV
1-50 ng/µL: 10 % CV

* Determined by analyzing the respective ladder as sample.
** Accuracy of software ladder ±20 %.
*** Measured against the Agilent 2200 TapeStation system.

15 % CV

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Agilent 2019

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