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The VSK-VMK004 DNA library preparation kit
The VolTRAX V2b provides an automated process for preparing a DNA library. The chemistry of the library preparation is the same as for manual library preparation for a MinION or GridION sequencing experiment.
Below is an overview of the library preparation stages using a VolTRAX V2b and the VolTRAX Multiplex Kit (VSK-VMK004).
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VolTRAX Multiplex Kit (VSK-VMK004) contents
Name Acronym Number of vials Cap colour Fill volume per vial (µl) VolTRAX Buffer Solution VBS 1 Orange 100 EDTA EDTA 1 Clear 20 VX AMPure XP Beads VXP 1 Pink 45 VX S Fragment Buffer VSF 1 White 30 VX Rapid Adapter T VRA T 1 Green 25 VX Elution Buffer VEB 1 Black 65 VX Running Buffer VRB 1 Red 250 VX BC Frag Mix 01-10 VB01-10 10 Amber 20 Flush Buffer FB 3 Blue 1,170 Flush Tether FLT 1 Purple 200 -
VolTRAX Multiplex Kit barcode sequences
Component Sequence VB01 AAGAAAGTTGTCGGTGTCTTTGTG VB02 TCGATTCCGTTTGTAGTCGTCTGT VB03 GAGTCTTGTGTCCCAGTTACCAGG VB04 TTCGGATTCTATCGTGTTTCCCTA VB05 CTTGTCCAGGGTTTGTGTAACCTT VB06 TTCTCGCAAAGGCAGAAAGTAGTC VB07 GTGTTACCGTGGGAATGAATCCTT VB08 TTCAGGGAACAAACCAAGTTACGT VB09 AACTAGGCACAGCGAGTCTTGGTT VB10 AAGCGTTGAAACCTTTGTCCTCTC -
VolTRAX Multiplex Kit chemistry
The VSK-VMK004 library preparation protocol uses our rapid chemistry, with a bead-based clean-up step. The kit contains ten barcodes for multiplexing up to ten samples in one library preparation.
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An overview of the VolTRAX Multiplex Kit protocol
1. Loading a protocol
- Use the protocol selector in the UI to load the VSK-VMK004 protocol.
- When prompted, connect your VolTRAX V2b device, press Continue and await connection.
2. System QC - Cartridge Test
- When prompted, connect a cartridge to the VolTRAX device and press Continue to proceed to the Cartridge Test.
3. Loading the Priming Fluid and reagents into the cartridge
- Prime the cartridge by twisting the oil container and removing the foil tab.
- The UI will recommend the quality, concentration, buffer and Absorbance (A) readings for your input DNA. These are stated at the stage of 'Reagent Loading' later in this protocol.
- Follow the instructions on the equipment required and pipetting technique.
- Load the appropriate reagents and extracted DNA into the ports as instructed by the UI.
- The reagents will be manipulated by the VolTRAX V2b to prepare the library for nanopore sequencing.
4. Extracting the library from the cartridge
- Follow the extraction instructions, using UI as a visual guide, to slowly extract the library from the cartridge, then transfer it into an Eppendorf tube.
- Prepare the library for loading onto the flow cell.