Oxford nanopore technology does not experience any fragment size limitation, not even size bias in the sense of preferential loading of short fragments or similar. On the other hand, it is very sensitive to any kind of inhibitors. Inhibition of nanopore activity might significantly decrease sequencing output. So carefully check samples for presence of inhibitors.

If you are not able to get rid of certain amount of inhibitors, we can construct low input library using few PCR cycles, inhibitors would be diluted and additionally purified. We recommend you freeze samples directly after isolation and quality control and avoid repeated thaw/freeze cycles.

For preparation of high-molecular weight gDNA please follow our recommendations for PacBio sequencing.

Note: For metagenomic analyzes based on the sequencing of long amplicons (e.g. 16S), gDNA can be prepared in a conventional manner as in the case of Illumina technology. It is important that the DNA is free of inhibitors of the PCR reaction.