The industry standard for sequencing long-read fragments is PacBio’s Single Molecule, Real-Time Sequencing technology. The PacBio systems allow you to directly sequence single molecules of DNA and achieve long sequencing reads with uniform coverage with no bias across GC content-rich regions.
Pacbio’s Single Molecule, Real-Time (SMRT) Sequencing technology consistently produces some of the longest average read lengths available in the industry (average > 10,000 bp, some reads > 60,000 bp). This gives users the flexibility to sequence a range of fragment sizes from amplicons to large insert libraries with lengths in excess of 30kb.
Advantages of Long Sequence Reads
Long Sequence Reads improve mappability for resequencing and simplify de novo assembly. They also allow for phasing and structural variation detection. SMRT sequencing enables users to catalog full-length isoforms in their transcriptomes and identify methylation signatures.
The long reads generated by PacBio SMRT sequencing technology can be used to generate long-read scaffolds which can be used to align short-read fragments for increased coverage and error correction. With the Iso-Seq application, users are able to sequence full-length transcripts to identify isoform expression patterns and integrate that information with their existing short-read RNASeq data.
Library preparation for PacBio requires high-quality, High Molecular Weight (HMW) DNA, and is priced based on how many SMRT cells are required. Because each library prep will only provide enough DNA for a certain number of SMRT cells, and longer fragment libraries require more library per SMRT cell, the cost of library prep is dependent on the type of project and depth of sequencing needed.
RTLGenomics can create libraries or can run user-created libraries. Contact us for more information on sample requirements (generally assume you will need 5-10ug minimum of HMW DNA).