Difference between revisions of "BUGSS:Build-a-BUG:2-3"

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=== Polymerase chain reaction (PCR) protocol ===
 
=== Polymerase chain reaction (PCR) protocol ===
* PCR mix
+
* Mixture
 
** PCR master mix - 45.0 μL
 
** PCR master mix - 45.0 μL
 
*** Nuclease-free water -35.6 μL
 
*** Nuclease-free water -35.6 μL
Line 23: Line 23:
 
*** ''Taq'' DNA polymerase - 0.5 μL
 
*** ''Taq'' DNA polymerase - 0.5 μL
 
** Template DNA (3002) - 5.0 μL
 
** Template DNA (3002) - 5.0 μL
 +
* Thermal cycler program - 30 cycles
 +
** Melt at 95°C for ? sec
 +
** Anneal at ?°C for ? sec
 +
** Extend at 72°C for ? sec
  
 
=== Materials ===
 
=== Materials ===

Revision as of 12:23, 18 March 2013

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Build-a-BUG: Series 2, Session 3

Saturday, March 9, 2013
Noon to 4:00 PM

This is the third Build-A-BUG workshop in a series of five on yeast. You will learn more about molecular cloning and continue to hone your basic lab techniques, while continuing either your own or somebody else's mating type detector project.

Background reading

Polymerase chain reaction (PCR) protocol

  • Mixture
    • PCR master mix - 45.0 μL
      • Nuclease-free water -35.6 μL
      • 10X PCR buffer - 5.0 μL
      • Forward primer (VF2) - 1.6 μL
      • Reverse primer (VR) - 1.3 μL
      • Deoxynucleotide Triphosphate (dNTP) Mix - 1.0 μL
      • Taq DNA polymerase - 0.5 μL
    • Template DNA (3002) - 5.0 μL
  • Thermal cycler program - 30 cycles
    • Melt at 95°C for ? sec
    • Anneal at ?°C for ? sec
    • Extend at 72°C for ? sec

Materials