PacBio Library Prep

Amplicon Sequencing 

Amplicons 500–10,000 bp can be prepped for sequencing on Revio using a standard SMRTbell library prep. We offer 96 SMRTbell barcodes for multiplexing, but recommend that you incorporate barcodes during PCR to reduce overall cost. Requires 100-500ng of pooled amplicons per SMRTcell in 25-50ul of water or low-EDTA buffer.  

Kinnex 

Kinnex library prep concatenates together individual inserts to increase the number of reads yielded from one SMRTcell. 

Full-length 16S

12-mer concatenation of full-length 16S amplicons. We have 96 full-length 16S barcodes and 4 Kinnex SMRTbell barcodes available, allowing up to 384 samples to run on one SMRTcell. Requires >5 ng of DNA per sample suspended in 25ul of water or low-EDTA buffer.  

If you plan to prepare your own 16S amplicons, please contact us to coordinate downstream compatibility. Submit >50 ng of purified, multiplexed amplicons.

Full-length 10X cDNA 

16-mer concatenation of 10X cDNA product. Requires >20ng cDNA in 20ul of elution buffer.  

Full-length RNA  

8-mer concatenation of IsoSeq full-length RNA. We have 12 IsoSeq barcodes and 4 Kinnex SMRTbell barcodes available, allowing up to 48 samples to run on one SMRTcell. Requires >500ng of high-quality RNA (RIN/RQN>7) per library suspended in 25ul of RNase-free water.

If you plan to prepare your own cDNA, please contact us to coordinate downstream compatibility. Submit >75 ng of purified, multiplexed cDNA.

Whole-Genome Sequencing 

PCR-free library prep of native DNA, generating 10–30 kb HiFi reads with 5mC calls. Up to 96 WGS libraries can be pooled for sequencing. Requires 2-5ug of HMW DNA (>50% of the gDNA must be >30kb in length) in 50-200ul of nuclease-free water or buffer. DNA should be free of visible particulates, dust/lint, precipitates, colors, or cloudiness, with Nanodrop 260/280 ratio >1.8 and 260/230 2.0-2.2.

PacBio library prep and sequencing success is highly dependent on the quality of input DNA for library prep. Contact us to discuss extraction optimization, cleanup, and/or size selection to help get your samples within QC requirements. 

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If you don’t see the right prep or aren’t sure what you need, email us at genomics@uoregon.edu.