PacBio Low Input Sequencing. Lambdavirus lambda

The PacBio SMRT sequencing technology provides several unique capabilities that differentiate it from other Next-Generation Sequencing technologies, such as very long read lengths and the ability to identify base modifications. However, unlike other NGS technologies, the PacBio library preparation process does not employ any amplification steps, so the input requirements (in terms of DNA mass) are much higher. This requirement can become an impediment to many sequencing experiments where the DNA is limited. Our paper describes a simple, cost-effective solution to the challenge of input requirements for PacBio library construction, while also describing ways to optimize sequencing conditions and re-use the libraries to obtain additional sequence data. We demonstrate that the method can work for both short (2kb) and long (20kb) targets and we believe this method can enable many sequencing projects that were previously impossible due to input limitations.