IGEM:UC Berkeley/2006/PCRPrep

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Line 5: Line 5:
#*0.5uL dNTPs (10uM)
#*0.5uL dNTPs (10uM)
#*0.1uL Taq
#*0.1uL Taq
-
#*0.5uL Forward primer
+
#*0.5uL Forward primer (10uM)
-
#*0.5uL Reverse primer
+
#*0.5uL Reverse primer (10uM)
#Add mixture to PCR tube.
#Add mixture to PCR tube.
#Pick colony and add to tube and then streak an "X" on a plate while making a map of which colony is where.
#Pick colony and add to tube and then streak an "X" on a plate while making a map of which colony is where.

Revision as of 14:52, 28 June 2006

Colony PCR

  1. Create PCR mixture.
    • 21.8uL
    • 2.5uL or 10X thermol pol buffer
    • 0.5uL dNTPs (10uM)
    • 0.1uL Taq
    • 0.5uL Forward primer (10uM)
    • 0.5uL Reverse primer (10uM)
  2. Add mixture to PCR tube.
  3. Pick colony and add to tube and then streak an "X" on a plate while making a map of which colony is where.


PCR Prep

  1. 38.5 ul water
  2. 5 ul buffer2
  3. 5 ul dNTP
  4. 1 ul oligo1
  5. 1 ul oligo2
  6. 1 ul DNA template
  7. .75 ul polymerase

50 ul total volume

Combine in PCR tube.

Notes:

To make 1ul oligo at correct concentration, take 1ul oligo and 9ul water, combine, transfer 1ul of solution to PCR tube

Keep polymerase on ice at all times

39 ul water can be used in place of 38.5ul water

-DLK_062106

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