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Sequence Improvement

Long-Read Sequencing Technology

This lecture on Long-Read Sequencing Technology was created by Katie Sandlin for the 2024 National GEP Faculty Workshop. The lecture covers four major topics: 1) Review of RNA-Seq Short-read Sequencing, 2) Overview and benefits of Long-read Sequencing, 3) Oxford Nanopore Sequencing, and 4) Pacific Biosciences Sequencing. Each slide is scripted in the Notes section for reference.

Introduction to UNIX Exercise

A simple exercise on using simple Unix commands to navigate to different directories and manipulate files.

Common Finishing Errors

This PowerPoint presentation describes the common errors observed in student finishing projects.

Reconcile Sequence Improvement Projects

All GEP projects are completed at least twice independently by GEP students. This document describes how to check two or more submissions of a finishing project for congruence. Ordinarily this is done centrally at Washington University, but in some cases may be of interest at a given school.

Dye Chemistry Characteristics

This document illustrates some of the advantages and potential problems with the BigDye and dGTP chemistries.

Fulton Lecture Notes

The lecture notes from a presentation by Bob Fulton point out some of the more advanced features of Consed, the base caller phred, and the assembler phrap. This document should be read after students have some experience using Consed but before they begin working on their fosmids.

Restriction Digests: A Tool to Check Your Findings

One of the main tools used to verify the correctness of a genome assembly is comparison of an in-silico restriction digest to the real restriction digest data generated by the WU Genome Center. This document also covers how students can use the restriction digest data early in the assembly process to determine the number of copies of repeats in a given cluster or to estimate gap sizes.

Finishing Lab Practice Using a Mouse Contig

Students can practice using Consed by working on a mouse contig. This exercise poses various challenges that students may encounter when working on their own projects.

A Complex Drosophila Fosmid

This fosmid from Drosophila virilis assembles into three contigs (a yellow clone). In this exercise, students must generate a final assembly by closing a gap, dealing with a mis-assembly, and improving low quality regions. Snapshots of the different stages of the assembly are stored as separate ace files.