Fluidigm Access Array - Thermal-Cycle the IFC; Harvest the IFC

Access Array System for the Ion Torrent PGM Sequencing System: User Guide
Chapter 5: Bidirectional Amplicon Tagging on the LP 48.48 IFC
Thermal-Cycle the IFC
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Thermal-Cycle the IFC
IMPORTANT If you are using the Access Array reagents with the LP 48.48 IFC, use only the
AA 48x48 Standard v1 or AA48v1 thermal protocols specified in step 8. Do not use the LP-
48x48 protocol with Access Array reagents.
Place the IFC onto the FC1 cycler and start PCR by selecting the protocol AA 48x48
Standard v1. Refer to the FC1™ Cycler User Guide (PN 100-1279).
For stand-alone thermal cycler, select protocol AA48v1.
NOTE The protocol as programmed into the Fluidigm stand-alone thermal cycler takes
undershooting and overshooting of target temperatures into consideration and is therefore
not identical to the PCR protocol for the FC1 cycler. Contact Technical Support if you need
assistance in programming your cycler.
Harvest the IFC
1 After the PCR has finished, move the IFC into the Post-PCR lab for harvesting.
2 Remove the remaining fluids from the H1–H4 wells.
3 Pipet 600 L of fresh 1X Access Array Harvest Solution into each of the H1–H4 wells. (Do
not use Hydration Reagent v2 here.)
NOTE If you are working with the LP 48.48 IFC or the Access Array 48.48 IFC, use 1X
Access Array Harvest Solution in all four wells (H1-H4).
4 Pipet 2 L of 1X Access Array Harvest Solution into each of the sample inlets on the IFC.
5 Load the IFC into the Post-PCR IFC Controller AX located in the Post-PCR lab.
6 Press Eject to move the tray out of the IFC Controller AX.
7 Place the IFC onto the tray by aligning the notched corner of the IFC to the A1 mark.
8 Press Load Chip to register the barcode of the IFC and activate the script selection.
9 Harvest the LP 48.48 IFC for Access Array. Select Harvest (155x) and Run Script.
NOTE For Access Array 48.48 IFCs, select Harvest v7 (151x) and Run Script. Harvest v7
(151x) is a script update from Harvest v5 (151x) for Access Array 48.48 IFCs and can be
downloaded from fluidigm.com/software. Contact Technical Support for assistance.
10 Once the script is complete, press Eject to remove the IFC.
11 Label a 96-well plate with the IFC barcode.
12 Carefully transfer 10 L of harvested PCR products from each of the sample inlets into
columns 1
–6 of a 96-well PCR plate, using an 8-channel pipette.
IMPORTANT Transfer PCR products from the IFC to the 96-well plate in the same order
as the IFC was loaded.