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 81
Option 2: Analysis using the Agilent 2200 TapeStation and High Sensitivity D1000
ScreenTape
Use a High Sensitivity D1000 ScreenTape (p/n 5067-5584) and reagent kit
(p/n 5067-5585) to analyze the amplified indexed DNA. For more
information to do this step, see the Agilent 2200 TapeStation User
Manual at www.genomics.agilent.com.
1 Prepare the TapeStation samples as instructed in the Agilent 2200
TapeStation User Manual. Use 2 µL of each indexed DNA sample
diluted with 2 µL of High Sensitivity D1000 sample buffer for the
analysis.
2 Load the sample plate or tube strips from step 1, the High Sensitivity
D1000 ScreenTape, and loading tips into the 2200 TapeStation as
instructed in the Agilent 2200 TapeStation User Manual. Start the run.
3 Verify that the electropherogram shows the peak of DNA fragment size
positioned between 250 and 350 bp. A sample electropherogram is
shown in Figure 10.
4 Measure the concentration of each library by integrating under the
entire peak.
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
overnight or at –20°C for up to one month.
CAUTION
Make sure that you thoroughly mix the combined DNA and sample buffer on a vortex
mixer for 5 seconds for accurate quantitation.