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- l Snap the tube strip holder with the Gel Bead strip into a 10x Vortex Adapter. Vortex 30 sec . |
- l Centrifuge the Gel Bead strip for ~5 sec after removing from the holder. Confirm there are no bubbles at the bottom of tubes and the liquid levels look even. Place Gel Bead strip back in the holder and secure the holder lid. |
- l If the required volume of beads cannot be recovered, place the pipette tips against the sidewalls and slowly dispense the Gel Beads back into the tubes. DO NOT introduce bubbles into the tubes and verify that the pipette tips contain no leftover Gel Beads. Withdraw the full volume of beads again by pipetting slowl... |
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## 10x Magnetic Separator B |
- l Images below are illustrative - actual appearance of magnetic separator may vary. Guidance applies to all 10x Magnetic Separators. |
- l Offers two positions of the magnets (high and low) relative to a tube, depending on its orientation. Flip the magnetic separator over to switch between high (magnet· High ) or low (magnet· Low ) positions. |
- l If using MicroAmp 8-Tube Strips, use only the high position (magnet· High ) throughout the protocol. |
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## Magnetic Bead Cleanup Steps |
- l During magnetic bead based cleanup steps that specify waiting 'until the solution clears', visually confirm clearing of solution before proceeding to the next step. See panel below for an example. |
- l The time needed for the solution to clear may vary based on specific step, reagents, volume of reagents etc. |
- l Images below are representative - actual color of magnetic separator may vary. Guidance applies to all 10x Magnetic Separators. |
## Visually Confirm Clearing of Magnetic Bead Solution |
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## SPRIselect Cleanup & Size Selection |
- l After aspirating the desired volume of SPRIselect reagent, examine the pipette tips before dispensing to ensure the correct volume is transferred. |
- l Pipette mix thoroughly as insufficient mixing of sample and SPRIselect reagent will lead to inconsistent results. |
- l Use fresh preparations of 80% Ethanol. |
## Tutorial - SPRIselect Reagent:DNA Sample Ratios |
SPRI beads selectively bind DNA according to the ratio of SPRIselect reagent (beads). |
Example Ratio: |
= Volume of SPRIselect reagent added to the sample |
= 50 μ l |
= 0.5X |
Volume of DNA sample |
100 μ l |
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## Enzymatic Fragmentation |
Ensure enzymatic fragmentation reactions are prepared on ice and then loaded into a thermal cycler pre-cooled to 4°C prior to initiating the Fragmentation, End Repair, and A-tailing incubation steps. |
## Sample Indices in Sample Index PCR |
- l Choose the appropriate sample index sets to ensure that no sample indices overlap in a multiplexed sequencing run. |
- l Verify and use the specified index plate only. DO NOT use the plates interchangeably. |
- l Each well in the Dual Index Plate contains a unique i7 and a unique i5 oligonucleotide. |
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