Editing BioMicroCenter:Cluster Visualization Protocol for Flowcell QC (section)

== Protocol ==

* Dilute 500uL of SYBR Green in 1500uL Wash Buffer in a 2mL microcentrifiuge tube labeled as #8 and load onto the Cluster Station

* Manually prime solution 8x10 and then manually pump the dilute SYBR using the following settings:

#Reagent: 8
#Flowrate: 30
#Volume 150

* Remove the Flowcell from the cluster station for observation on the Fluorescent microscope.  Use the GFP filter set, blue light. Neutral Denstity filters can be used to help prevent photobleaching

'''If amplification was successful and clusters were observed continue with protocol.  If clusters were not visualized re-cluster samples on a new Flowcell'''

* Place Flowcell back onto Cluster Station

* Label a 50mL tube as #10 and fill with about 5mL Wash Buffer.  Load #10 onto the Cluster station.

* Manually pump #10 using the following settings:

#Reagent: 10
#Flowrate: 30
#Volume: 200

* Label a 50mL tube as #12 and fill with about 5mL Storage Buffer. Load #12 onto the Cluster Station.

* Manually pump #12 using the following settings:

#Reagent: 10
#Flowrate: 30
#Volume: 200

* Remove Flowcell and store at 4C until ready for use

* Replace #8 with 1.5mL screw cap tube filled with water

* Remove amplification manifold and replace with wash bridge

* Was line 8 by manually pumping water using the following settings:

#Reagent: 8
#Flowrate: 30
#Volume: 150