454 pyrosequencing (Drosophilidae)

Knowledge graph centered on 454 pyrosequencing (Drosophilidae) with 33 nodes and 113 connections. Top connected: Ligusticum porteri, Lepidoptera, Arabidopsis, Boechera stricta, Brassica.

Method synopsis

454 GS FLX Titanium sequencing of normalized cDNA libraries to generate reference transcriptome assembly for gene discovery and annotation.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Genes involved in the evolution of herbivory by a leaf-mining drosophilid fly (2012), Genome Biology and Evolution. Implementations in other papers (listed below) may differ.

1. 454 transcriptome sequencing
   454 GS FLX library preparation followed previously published protocols. cDNAs were normalized and size selected for average size of 400-800 bp. 454 GS FLX Titanium-specific adapters were ligated to DNA ends and polished. Libraries were sequenced on 454 GSFLX flow cell using titanium chemistry at University of Illinois.
   Quantities: 400-800 bp average fragment size, single 454 GSFLX flow cellDuration: Not specifiedConditions: University of Illinois sequencing facility
   Equipment: 454 GS FLX Titanium sequencer, 454 GS FLX Titanium-specific adapters

Typical Equipment

• 454 GS FLX Titanium sequencer
• 454 GS FLX Titanium-specific adapters

Output Measurements

• DNA sequence reads
• transcriptome contigs

Papers Using This Protocol (1)

Species Studied With This Protocol