Article Title: Lane-by-lane sequencing using Illumina’s Genome Analyzer II
Figure Lengend Snippet: Setting up lane-by-lane sequencing on an Illumina GAIIx a) cBOT cluster plate set up to cluster lanes 5–8. Lanes 1–4 have been removed using a razor blade and replaced with vials containing HSB. b) The TileSelection section of an XML recipe file for 4-lane sequencing of the flowcell clustered in a. Only lanes 5–8 will be imaged. c) Disconnecting the syringe-pumps. Once the manifold housing is removed, the plunger heads can be unscrewed from the motorized crossbar. Here, pumping has been inactivated on lanes 1 and 2. d) Pause-and-pulse reagent delivery method for reads longer than 40 nt. The left lane is active and the right lane is disconnected. Orange arrows represent flow of new reagent, green arrows represent movement of previous reagent, white circle represents a small air bubble remaining in the disconnected lane. i. During the initial pumping stage, reagent delivered from the inlet (bottom) may be contaminated by fluid from disconnected lanes as small bubbles expand due to negative pressure from the pump. ii. After pumping, a 5-second pause allows a small amount of the current reagent to back-fill through the inlet manifold as pressure equalizes in the system. iii. A 20 μL pulse then finishes delivery with homogenous reagent.
Article Snippet: Flowcells can be clustered on any desired number of lanes using the Illumina cBot, sequenced, stored, and then clustered again on remaining lanes without appreciable reduction in intensity, cluster density or accuracy of the run.
Techniques: Sequencing, Flow Cytometry