Technical data

5 Post-Capture Sample Processing for Multiplexed Sequencing

Step 3. Assess quality with the 2100 Bioanalyzer High Sensitivity DNA assay

68 SureSelect

XT2

Target Enrichment System for Illumina

Step 3. Assess quality with the 2100 Bioanalyzer High

Sensitivity DNA assay

You can use Agilent’s 2200 TapeStation for rapid analysis of multiple samples at this step.

For analysis of captured DNA, use the High-Sensitivity D1000 ScreenTape (p/n 5067-5584)

and associated reagents. See the 2200 TapeStation and High-Sensitivity D1000 ScreenTape

protocols for information on sample preparation and data analysis.

Use a High Sensitivity DNA Assay kit to assess sample quality and

quantity using the 2100 Bioanalyzer. See the High Sensitivity DNA Kit

Guide at www.genomics.agilent.com, for more information on doing this

step.

1 Check that the 2100 Bioanalyzer electrodes have been cleaned as

instructed in the reagent kit guide.

2 Open the 2100 Expert Software (version B.02.07 or higher required to

run the High Sensitivity Kit), turn on the 2100 Bioanalyzer, and check

communication.

3 Prepare the chip, samples and ladder as instructed in the reagent kit

guide, using 1 µL of diluted captured library samples for the analysis.

4 Load the prepared chip into the 2100 Bioanalyzer and start the run

within five minutes after preparation.

5 Within the instrument context, choose the High Sensitivity DNA assay

from the drop down list.

6 Start the run. Enter sample names and comments in the Data and

Assay context.

7 Verify the results. Check that the electropherogram shows a distribution

with a fragment size peak between approximately 275 to 300 bp. A

sample electropherogram is shown in Figure 6.

8 Determine the concentration of each captured indexed library pool by

integration under the peak in the electropherogram.

NOTE

Prior to Bioanalyzer analysis, dilute each amplified captured library sample ten- fold in TE

buffer, as described in step 18 of the previous section.