Information
This kit is recommended for users who:
- Want to optimise their sequencing experiment for throughput
- Require control over read length
- Would like to utilise upstream processes such as size selection or whole genome amplification
Please note that to use this kit, you will need to purchase additional 3rd party reagents: see the "3rd party materials" tab for more detail.
The Q-Line Ligation Sequencing Kit offers a flexible method of preparing sequencing libraries from dsDNA (e.g. gDNA, cDNA or amplicons). The library preparation method is straightforward: DNA ends are repaired and dA-tailed using the NEBNext Companion Module v2, and then sequencing adapters, supplied in the kit, are ligated onto the prepared ends.
The Q-Line Ligation Sequencing Kit features:
| Feature | Property |
|---|
| Preparation time | 65 minutes |
| Input requirement | 1,000 ng for samples >10 kb or 200 ng for samples ≤10 kb |
| PCR Required | No |
| Fragmentation | Optional |
| Read length | Equal to fragment length |
| Kit chemistry | Kit 14 |
| Pack size | 6 reactions |
| Stability | Store at –30°C to –20°C until the labelled expiration date.
Q reagents are supplied with a minimum of 3 months shelf life remaining at the time of delivery.
Expiration dates are displayed in DD-MMM-YYYY format. |
If your experiment requires long reads, it is recommended to start with full-length gDNA, and fragmentation/shearing is neither advised nor required. Please note: if long reads are required, then long fragments must be present in the starting material. To determine purity, we suggest using the NanoDrop to measure the A260/280 and A260/230 ratios and we recommend that the sample should meet the following criteria:
- A260/280 = 1.8
- A260/230 = 2.0–2.2
Further considerations:
Starting with lower amounts of input material, or impure samples, can affect library preparation efficiency and can reduce sequencing throughput. PCR can be used to generate more input material in cases where sample amount is limiting. Also, where sample purity is compromised and library preparation efficiency may be reduced, PCR will select for molecules of higher quality, and generate more of them. The original impurity is then lost or diluted.
Shipping and logistics:
Bulk order options are available. All orders can be split for delivery to best suit your needs.
Flow cells and kits are shipped together at 2°C to 8°C. The packaging is designed to protect the flow cells from freezing.
Workflow
The Q-Line Ligation Sequencing Kit V14 offers a flexible method of preparing sequencing libraries from dsDNA (e.g. gDNA, cDNA or amplicons). The library preparation method is straightforward: DNA ends are FFPE repaired and end-prepped/dA-tailed using the NEBNext Companion Module v2, and then sequencing adapters, supplied in the kit, are ligated onto the prepared ends.
What's in the box
The Q-Line Ligation Sequencing Kit V14 contains sufficient reagents to generate six sequencing libraries.
| Name | Abbreviation | Cap colour | Quantity | Minimum fill volume per container (µl) |
|---|
| Ligation Adapter | LA | Green | 1 | 30 |
| AMPure XP Beads | AXP | Amber | 1 | 600 |
| Ligation Buffer | LNB | White | 1 | 150 |
| L Fragment Buffer | LFB | Orange | 2 | 1,500 |
| S Fragment Buffer | SFB | Clear | 2 | 1,500 |
| Sequencing Buffer | SB | Red | 1 | 225 |
| Elution Buffer | EB | Black | 1 | 90 |
| Library Beads | LIB | Pink | 1 | 153 |
| Flow Cell Flush | FCF | N/A | 1 | 7,020 |
| Flow Cell Tether | FCT | Purple | 1 | 180 |
Note: The AMPure XP reagent manufactured by Beckman Coulter Inc. can be stored at –30°C to –20°C with the kit, without detriment to reagent stability.
Compatibility
The Q-Line Ligation Sequencing Kit V14 can be used together with:
Kits
- Q-Line Control Expansion Kit (Q-EXP-CTL001)
- Q-Line Flow Cell Wash Kit (Q-EXP-WSH004)
Flow cells
- Q-Line MinION Flow Cell R10 (Q-FLO-MIN114)
Devices
- GridION Q sequencing device (Q-GRD-MK1)
