Protocols in Production Sequencing

Sanger

Libraries

Large Clone Isolation Protocol
DNA Quantification Protocol

Library Creation

• General 3 kb Protocol
  Vector: pUC18; restriction site: CCCGGG; sequence
• Mid-size 8-10 kb Protocol
  Vector: pMCL200; restriction site: GATATC; sequence (Gene 162, 157-158, 1995.)
• Fosmid 40 kb Protocol
  Vector: pCC1Fos; restriction site: CACGTG; sequence

Library Plating Protocol
Q-Pix Colony Picker Protocol
Fosmid DNA Inoculation Protocol

Sequencing Preparation

Rolling Circle Amplification Protocol using TempliPhi
Sequencing Chemistry:

Plasmid
Fosmid

• Big Dye 3.1 Sequencing of Fosmid DNA Protocol
• Big Dye 3.1 Cycle Sequencing Protocol

BET Cleanup Protocol

Sequencers

ABI3730xl Users Protocol

454 Titanium

Libraries

• Paired-end
• Standard Shotgun

Sequencing Preparation

• 454 Titanium emPCR Bulk Protocol

Sequencers

• 454 titanium Sequencing Run Protocol

Illumina

Libraries

• Std Library Creation Protocol
• Normalized Std library Creation Protocol

Sequencing Preparation

• Cluster Generation Protocol

Sequencers

• Genome Analyzer 2 Run Protocol