This guide describes an optimized procedure for preparing your DNA template for sequencing on the PacBio System. This Blunt-End Liga-tion protocol, is suitable for all libraries (greater than 250 bp amplicons) and offers a total preparation time of approximately four hours. SMRTbellTM Templates.

---

Use of these protocols is offered to those customers who understand and accept the associated terms and conditions and wish to take advantage of their potential to help prepare samples for analysis using the PacBio® System.

---

Use of these protocols is offered to those customers who understand and accept the associated terms and conditions and wish to take advantage of their potential to help prepare samples for analysis using the PacBio® system.

---

This Bulletin provides recommendations and tips for preparing 20 kb SMRTbell templates using the BluePippinTM size-selection method using size-selection cutoffs between 6 kb and 10 kb. Once you have read this User Bulletin, proceed to the Procedure & Checklist - 20 kb Template Preparation and Sequencing for pre-paring your 20 kb SMRTbell templates.

---

Guidelines for Preparing 20 kb SMRTbell™ Templates. This procedure can be used to prepare size-selected libraries from 5 μg of sheared and concentrated DNA using the BluePippin Size-Selection system.

---

Shared Protocol - Extracting DNA using Phenol-Chloroform. Please note: the shared protocols described herein may not have been validated by Pacific Biosciences and are provided as-is and without any warranty.

---

Combining xGen Lockdown probes with SMRT Sequencing provides a method for completely sequenced candidate genes and their corresponding full-length transcripts. This method enables: Detection of a broad range of genomic variants, from SNPs to multi-kilobase insertions and deletions.

---

Supported applications are sequencing of one species per sample or loci. Examples of supported applications include: Confirmation of SNPs, resequencing, most Long Amplicon Analysis (LAA) applications, and Sanger sequencing replacement. An exception is HLA typing, which may have 2 species per loci.

---

This SMRTbell library cleanup protocol involves a) binding annealed PacBio DNA SMRTbell libraries to magnetic beads, b) washing the bound annealed DNA SMRTbell templates to remove potential contaminants, and c) eluting the purified, annealed DNA SMRTbell templates from the magnetic beads.

---

It describes how to use a high-salt low-ethanol percentage wash to remove polysaccharides before DNA is precipitated from the solution. Prepare a buffer of 1 M NaCl and 2 mM EDTA. Label tube as Buffer A and set aside. Note that 500 μL of buffer will need to be prepared for each sample.

---