OP-HELISCOPE-sequencing-v1.0

Protocol: OP-HELISCOPE-sequencing-v1.0

Author: Kawashima, Tsugumi

Created: Apr 11th, 2011

Update: Apr 11th, 2011

Parameters: machine, run, flowcell, channel

Description:

Step 1. PolyA tailing of the DNA

Perform Poly-A Tailing Reaction and 3’ End Blocking Reaction according to the instructions found in the Helicos(TM) Chromatin Immunoprecipitation-Sequencing (ChIP-Seq)　Sample Preparation Protocol - Direct Tailing Procedure (Version 121310) [1].

Step 2. Sample loading on HeliScope flow cells

a. Prepare 20 μl of each sample in Hybridization Buffer according to the instructions found in the HeliScope(TM) Sample Loader Manual (LB-016), Step 1b-2 [2].

b. Transfer and hybridize the sample to the flow cells according to the instructions found in the HeliScope(TM) Low-Volume Sample Loading Protocol (LB-017 Rev. 01) [3].

Step3. Sequencing on HeliScope Single Molecule Sequencer

Transport the flow cells [flowcell] containing the sample on channel [channel] to the HeliScope Single Molecule Sequencer with using machine [machine] and run the sequencing process with run [run] according to the manufacturer’s manual, Helicos BioSciences Corporation Helicos(TM) Genetic Analysis System (LB-001 Rev. 04).

References/Notes:

[1] ‘Helicos(TM) Chromatin Immunoprecipitation-Sequencing (ChIP-Seq) Sample Preparation Protocol - Direct Tailing Procedure (Version 121310)’ [2] ‘HeliScope(TM) Sample Loader Manual (LB-016)’ [3] ‘Helicos(TM) Low-Volume Sample Loading Protocol (LB-017 Rev. 01)’ [4] ‘Helicos BioSciences Corporation Helicos(TM) Genetic Analysis System (LB-001 Rev. 04)’