AlanFung:Protocil/Cluster Station: Difference between revisions
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===perform linearization blocking and primer hybridization=== | ===perform linearization blocking and primer hybridization=== | ||
*load reagents in the following positions as indicated | *load reagents in the following positions as indicated | ||
single | *single-read recipe-load position 3,4,7,12,15,17 | ||
*paired-end recipe-load position 3,4,7,12,14,17 | |||
*click ok to resume the recipe and follow the prompt to proceed to linearization blocking and primer hybridization | |||
*load flow cell and attach the amplification manifold | |||
*click ok to proceed and follow the prompt to check for proper flow | |||
*as the process starts check for correct fluid flow through all eight lanes and the lines of the amplification manifold | |||
when the flow is regular in all lanes proceed with the protocol | |||
*click ok to proceed | |||
*wait for the message to let you know that the flow cell is ready for sequencing | |||
*remove the flow cell from the cluster station | |||
DO NOT STORE THE FLOW CELL AT THIS POINT FOR LONG PERIODS OF TIME. SEQUENCING MUST BE PERFORMED ON THE FLOW CELL WITHIN 4 HOURS | |||
* |
Revision as of 14:41, 21 July 2009
Performing linearization, blocking , and primer hybridization
wash the cluster station
- select file|open recipe
- open the standard protocol recipe and click run
- install the washing bridge and load containers filled with water in the following positions
*single-read recipe-load position 3,4,7,12,15,17 *paired-end recipe-load position 3,4,7,12,14,17
- click ok to proceed and follow the prompts when the wash is complete
- remove water from the following positions
*single-read recipe-load position 3,4,7,12,15,17 *paired-end recipe-load position 3,4,7,12,14,17
- click ok and follow the prompts when the air gap has been primed
loading reagents
best practices always remove and replace reagents one tube at a time wear gloves at all times do not touch reagents with bare hands connect the tubes by holding the caps stationary while you twist the tubes into place to prevent crimping and twisting of the lines
perform linearization blocking and primer hybridization
- load reagents in the following positions as indicated
*single-read recipe-load position 3,4,7,12,15,17 *paired-end recipe-load position 3,4,7,12,14,17
- click ok to resume the recipe and follow the prompt to proceed to linearization blocking and primer hybridization
- load flow cell and attach the amplification manifold
- click ok to proceed and follow the prompt to check for proper flow
- as the process starts check for correct fluid flow through all eight lanes and the lines of the amplification manifold
when the flow is regular in all lanes proceed with the protocol
- click ok to proceed
- wait for the message to let you know that the flow cell is ready for sequencing
- remove the flow cell from the cluster station
DO NOT STORE THE FLOW CELL AT THIS POINT FOR LONG PERIODS OF TIME. SEQUENCING MUST BE PERFORMED ON THE FLOW CELL WITHIN 4 HOURS