Access Array System for the Ion Torrent PGM Sequencing System: User Guide
Chapter 6: Post-PCR Amplicon Purification and Quantitation
PCR Product Library Quantitation Procedure
38
6
Vortex the tube and incubate at room temperature for 10 minutes.
7
Place the microtube onto a magnetic separator and allow it to sit for 1 minute.
8
Carefully pipet out the supernatant without disturbing the beads (remove as much liquid
as possible).
9
Add 180
μ
L of 70% ethanol and vortex for 10 seconds.
10
Place the microtube onto a magnetic separator and allow it to set for 1 minute.
11
Carefully pipet the supernatant without disturbing the beads.
12
Add 180
μ
L of 70% ethanol and vortex for 10 seconds.
13
Place the microtube onto a magnetic separator and allow it to set for 1 minute.
14
Carefully pipet out the supernatant without disturbing the beads.
15
Allow the beads to air dry for approximately 10 minutes by leaving the tube on the bench.
Make sure the tube is completely dry before proceeding.
16
Add 40
μ
L of DNA suspension buffer to the microtube and vortex for 5 seconds.
17
Place the microtube onto a magnetic separator and allow it to set for one minute.
18
Carefully transfer the supernatant to a new 1.5 mL microtube.
PCR Product Library Quantitation Procedure
Use one of the following quantitation methods to quantify the PCR product library.
IMPORTANT
The Ion Library Quantification Kit (Ion Torrent, PN.4468802) is not compatible
with the library generated with Access Array Barcode Library for Ion
Torrent™ PGM™ Sequencer
- 96
(Fluidigm, PN 100-4911).
Agilent 2100 Bioanalyzer Quantification
1
Run 1
μ
L of the PCR product library on a Bioanalyzer DNA 1000 Chip following the
manufacturer’s instructions.
2
Define a region of interest in the electropherogram to determine the PCR product library
concentration.
a
Select the Region Table subtab on the bottom panel of the Electropherogram tab.
b
Right-click the electropherogram and select Add region. Define the region to cover
all of the PCR product library peaks.
