Technical data

Indexing and Sample Processing for Multiplexed Sequencing 5
Step 3. Assess indexed library DNA quantity and quality
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 79
Step 3. Assess indexed library DNA quantity and quality
Option 1: Analysis using the Agilent 2100 Bioanalyzer and High Sensitivity DNA
Assay
Use the Bioanalyzer High Sensitivity DNA Assay to analyze the amplified
indexed DNA. See the High Sensitivity DNA Kit Guide at
www.genomics.agilent.com for more information on doing this step.
1 Set up the 2100 Bioanalyzer as instructed in the reagent kit guide.
2 Prepare the chip, samples and ladder as instructed in the reagent kit
guide, using 1 µL of each sample for the analysis.
3 Load the prepared chip into the 2100 Bioanalyzer and start the run
within five minutes after preparation.
4 Verify that the electropherogram shows the peak of DNA fragment size
positioned between 250 and 350 bp. A sample electropherogram is
shown in Figure 9.
5 Measure the concentration of each library by integrating under the
entire peak. For accurate quantification, make sure that the
concentration falls within the linear range of the assay.
If you wish to more-precisely quantify the target enriched samples prior
to pooling, proceed to “Step 4. Quantify each index-tagged library by
QPCR (optional)” on page 83.
Otherwise, proceed to “Step 5. Pool samples for multiplexed
sequencing” on page 84.
Stopping Point If you do not continue to the next step, store the libraries at 4°C for up
to one week or at –20°C for longer periods.