Fragment DNA / Recombine and Fragment DNA
If running with PGx, this step creates a combined whole genome and target PGx library followed by a fragmentation step to enzymatically fragment the DNA. An endpoint fragmentation is used to prevent over-fragmentation.
If running without PGx, this step enzymatically fragments the DNA. An endpoint fragmentation is used to prevent over-fragmentation.
Preparation
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1.
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Prepare the following consumables: |
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FMS
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-25°C to -15°C
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Thaw at room temperature. Vortex or invert 10 times to mix, and then centrifuge at 280 x g for 1 minute.
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2.
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Prepare tubes of FMS according to instructions on the interface. |
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0.5
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1
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1
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2
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2
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4
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3
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6
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4
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8
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5
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10
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6
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12
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3.
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Preheat the heat block with the MIDI plate insert to 37°C. |
Procedure
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1.
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Remove the WGA plate from the Illumina Hybridization Oven after amplification is complete. |
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2.
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[PGx] Remove the PGx plate from 4°C storage after amplification is complete. |
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3.
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Centrifuge the WGA and PGx(PGx) plates at 280 × g for 1 minute. |
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4.
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Open ILASS on the Infinium Automated Pipetting System. |
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5.
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Select an ILASS workflow as follows: |
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a.
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If running with PGx, select Infinium Genotyping Assay with PGx, then Recombine and Fragment DNA. |
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b.
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If running without PGx, select Infinium Genotyping Assay, then Fragment DNA. |
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6.
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Select the Product Type drop-down, and then select your product type.
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7.
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In the Container Input Quantity box, enter the number of 96-well plates to be processed. Manually enter the value or set the value using +/-.
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ILASS displays the number of required materials based on number of plates.
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9.
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Set up the Infinium Automated Pipetting System worktable as shown on the interface and as follows: |
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a.
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If running with PGx, set according to number of PGx-WGA pair(s) and FMS tube(s) used in the run. |
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b.
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If running without PGx,
set according to number of WGA plate(s) and FMS tube(s) used in the run.
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10.
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Before loading the worktable, mix FMS by inverting 10 times and centrifuge at 280 x g for 1 minute.
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11.
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Unscrew FMS caps and load in the tube rack at the assigned location displayed on the interface. Load enough tubes to account for the number of sample plates. |
The FMS tube(s) must be placed in the innermost tube rack position first.
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12.
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Select Auto-Scan Item(s). |
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13.
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After successful scanning, the user interface displays a timer.
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15.
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Wait for the Infinium Automated Pipetting System to finish the run.
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16.
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Remove WGA plate(s) and PGx plate(s) (PGx) from the worktable.
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17.
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[PGx] Mark the WGA barcode checkbox when PGx transfer is complete.
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18.
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Apply cap mat to WGA plate(s).
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19.
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[PGx] Apply foil adhesive seal to the PGx plate(s) and store at -25°C to -15°C.
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20.
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Remove tubes from the worktable. Discard residual reagent volume from tubes. Discard tubes. |
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21.
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Vortex WGA plate(s) at 1600 rpm for 1 minute. |
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22.
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Centrifuge at 280 × g at room temperature for 1 minute. |
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23.
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Incubate the sealed plate(s) on the 37°C heat block for 1 hour. |
Leave the plate(s) in the 37°C heat block until you have completed preparation for the next step. Do not leave the plate(s) on the heat block for longer than 2 hours.
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24.
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Centrifuge the plate(s) at 280 × g for 1 minute to remove condensation from the seal before storage. |
