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Sample Preparation
Step 6. Assess library DNA quantity and quality
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Option 2: Analysis using the Agilent 2200 TapeStation and D1000 ScreenTape
Use a D1000 ScreenTape and associated reagent kit to analyze the
amplified libraries. For more information to do this step, see the Agilent
2200 TapeStation User Manual.
1 Seal the DNA sample plate using the PlateLoc Thermal Microplate
Sealer, with sealing settings of 165°C and 1.0 sec.
2 Vortex the plate to mix samples in each well, then centrifuge the plate
for 30 seconds to drive the well contents off the walls and plate seal.
3 Prepare the TapeStation samples as instructed in the Agilent 2200
TapeStation User Manual. Use 1 µL of each amplified library DNA
sample diluted with 3 µL of D1000 sample buffer for the analysis.
CA U T I O N
Make sure that you thoroughly mix the combined DNA and D1000 sample buffer on a
vortex mixer for 5 seconds for accurate quantitation.
4 Load the sample plate or tube strips from step 3, the D1000
ScreenTape, and loading tips into the 2200 TapeStation as instructed in
the Agilent 2200 TapeStation User Manual. Start the run.
5 For each sample, measure the concentration of the library (ng/µL) by
integrating under the peak at approximately 180 to 550 bp. A sample
electropherogram is shown in Figure 9.
Stopping Point
If you do not continue to the next step, seal the plate and store at –20°C.
Figure 9
Analysis of amplified library DNA using the Agilent 2200 TapeStation.
SureSelectXT Automated RNA Library Prep and Target Enrichment for Illumina Sequencing
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