OP-HELISCOPE-sequencing-v1.0
From FANTOM5_SSTAR
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.
References/Notes:
[1] ‘Helicos(TM) Chromatin Immunoprecipitation-Sequencing (ChIP-Seq) Sample Preparation Protocol - Direct Tailing Procedure (Version 121310)’
File:Helicos ChIP Seq Protocol.pdf
[2] ‘HeliScope(TM) Sample Loader Manual (LB-016)’
File:LB-016 01, Sample Loader Manual.pdf
[3] ‘Helicos(TM) Low-Volume Sample Loading Protocol (LB-017 Rev. 01)’
