Most NGS reagent kits capable of generating 110 nt reads (single-end) are suitable for digitalMLPA reactions. For the Illumina NextSeq, it is also possible to use a 75-cycle kit with custom primers (see below). We don't recommend using the Illumina MiSeq Reagent Kits v2, because they may affect certain probes leading to false results.
Recommended NGS reagent kits per sequencer:
| Instrument | Recommended Flow Cell / Reagent Kit | Catalogue Number |
|---|---|---|
| Illumina iSeq | Standard iSeq flow cell. | – |
| Illumina MiSeq | Illumina MiSeq Reagent Kit v3 (150 cycle). | MS-102-3001 |
| Illumina MiniSeq | Illumina MiniSeq High Output Kit (150 cycle). | FC-420-1002 |
| Illumina NextSeq 500/550 | Illumina NextSeq 500/550 Mid Output Kit v2.5 (150 cycle). | 20024904 |
| Illumina NextSeq High Output Kit (75 cycle).
Custom primers are required. |
20024906 | |
| Illumina HiSeq, NovaSeq, NextSeq 1000/2000 | Any flow cell capable of generating 110 nt or more sequences in the first read. | – |
| GeneMind FASTASeq S, FASTASeq 300, GenoLab M, SURFSeq 5000, SURFSeq Q | Any flow cell capable of generating 110 nt or more sequences in the first read. | – |
Background
digitalMLPA requires single-end reads of 110 nt. When preparing the NGS run, select single read as Read type and set the Read Lengths (Read 1) to 110 in the run settings. For Illumina sequencers, you can also use our digitalMLPA Library Prep Kit template.