AlanFung:Protocol/Genome Analyzer: Difference between revisions

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>Alan6017518
Line 12: Line 12:
  6          Cleavage Mix          50ml                50ml tube
  6          Cleavage Mix          50ml                50ml tube
  7          Cleavage Buffer        70ml                250ml bottle
  7          Cleavage Buffer        70ml                250ml bottle
==Scanning Mix==
Required Materials
Scanning Reagent
Scanning Buffer
Filter Unit
Scanning Mix Tube
*Add the Scanning Reagent vial to the Scanning Buffer on ice
*Rinse the empty vial into the Scanning Buffer three times with 1 ml Scanning Buffer
*Invert the Scanning Buffer several times to mix
*Leave on ice for 10 minutes
*Invert several times to mix. Check to ensure that the solids are completely dissolved
*Filter the Scanning Mix with the provided Scanning Mix Filter Unit
*Pour 50 ml of the Scanning Mix into the 50 ml Scanning Mix Tube
*If you are running more than 26 cycles on the Genome Analyzer, you must top off the Scanning Mix at the end of the first day of the sequencing run. The tube should be approximately one-third empty after running for six or seven hours
*Record the weight of the reagent in the lab tracking worksheet
*Leave the remaining solution in the Filter Unit at 4oC for later cycles
*Keep the Scanning Mix on ice until ready to load

Revision as of 18:27, 25 June 2009

Preparing Reagents for the Genome Analyzer

  • Genome analyzer Reagent Compartment & Reagent Position

File:Reagent Position.jpg

  • Position of each reagent in the Genome Analyzer
NOTE:LABEL ALL REAGENT CONTAINERS WITH THE APPROPRIATE NUMBER
Position    Reagent                Volume              Container
1           Incorporation Mix      Varies w/ cycle #   50ml amber tube
2           DI H2O                                     250ml bottle
3           Scanning Mix           50ml                50ml tube
4           High Salt Buffer                           250ml bottle
5           Incorporation buffer   150ml               250ml bottle
6           Cleavage Mix           50ml                50ml tube
7           Cleavage Buffer        70ml                250ml bottle

Scanning Mix

Required Materials
Scanning Reagent
Scanning Buffer
Filter Unit
Scanning Mix Tube
  • Add the Scanning Reagent vial to the Scanning Buffer on ice
  • Rinse the empty vial into the Scanning Buffer three times with 1 ml Scanning Buffer
  • Invert the Scanning Buffer several times to mix
  • Leave on ice for 10 minutes
  • Invert several times to mix. Check to ensure that the solids are completely dissolved
  • Filter the Scanning Mix with the provided Scanning Mix Filter Unit
  • Pour 50 ml of the Scanning Mix into the 50 ml Scanning Mix Tube
  • If you are running more than 26 cycles on the Genome Analyzer, you must top off the Scanning Mix at the end of the first day of the sequencing run. The tube should be approximately one-third empty after running for six or seven hours
  • Record the weight of the reagent in the lab tracking worksheet
  • Leave the remaining solution in the Filter Unit at 4oC for later cycles
  • Keep the Scanning Mix on ice until ready to load