Q&A with Genomics Core Facility Directors Pettersson also appeared on a panel of expert users in SMRT Sequencing as a Service – How to Bring Long-Read Technology to Your Core Lab. She shared tips for sample prep, instrument handling, and business planning, as well as some of the advantages of the Sequel IIe System.

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Adding a barcode to the PCR amplicon. This approach involves a two-step PCR reaction workflow. The internal primers for the first PCR are augmented at the 5’ end by universal sequences to the target-specific primers. The external primers contain the 16bp barcode at the 5’ end and the universal sequences.

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PacBio SampleNet – Shared Protocol Page 1 Please note: the shared protocols described herein may not have been validated by Pacific Biosciences and are provided as-is and ... it is the responsibility of the end user to perform the required validation. Guidelines for Using a Salt:Chloroform Wash to Clean Up gDNA . This protocol can be used to ...

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The Iso-Seq method can be used for a number of projects. Transcript identification and annotation allows users to detect alternatively spliced isoforms. Targeted sequencing can be used to show, for example, which isoforms are enriched in which tissues. Another application is normalization; the approach can be used to reduce the representation ...

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Adding a barcode to the PCR amplicon. This approach involves a two-step PCR reaction workflow. The internal primers for the first PCR are augmented at the 5’ end by universal sequences to the target-specific primers. The external primers contain the 16bp barcode at the 5’ end and the universal sequences.

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The SRE XS and SRE XL kits can be used for rapid high-pass size selection of high molecular weight (HMW) DNA samples. This method can significantly enhance mean read length by progressively depleting short DNA up to 10 kb (SRE XS kit) or 40 kb (SRE XL kit) in length (Figure 1). The kits use a centrifugation procedure similar to standard ethanol ...

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Here we describe SMRT-cappable-seq as the first experimental methodology to identify operon structures genome-wide in prokaryotes. It combines the isolation of full length primary transcripts with SMRT Sequencing. Applied to E. coli, SMRT-cappable-seq identifies a total of 2300 operons from which around 900 are novel.

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CUSTOMER SHARED METHODS This protocol was generously contributed by the Lowe Lab at Hopkins Marine Station, Pacific Grove, California Preparing Genomic DNA from Sperm This protocol can be used to prepare purified genomic DNA from sperm for sequencing on PacBio Systems. It is recommended to start with ~250 µl of whole sperm, which can generate

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What is Kinnex? The Kinnex kits are based on the MAS-Seq method, which concatenates smaller amplicons into larger fragment libraries for throughput increase. Kinnex kits increase throughput for full-length RNA, single-cell RNA, and 16S rRNA sequencing on PacBio long-read sequencing systems to enable large-scale studies at a resolution difficult ...

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Figure 1. Barcoded forward primer mix added across the rows A—H. Add the reverse 16S rRNA barcoded primer down the column of the 96-well PCR plate as shown in figure 2. For the 1st 96-well plate use the first 12 reverse primers, and for the 2nd 96-well plate use the last 12 reverse primers listed in the appendix. Figure 2.

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