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Worm PCR

Single Worm PCR


Lysis Buffer:

50 mM KCl

10 mM Tris pH 8.3

2.5 mM MgCl2

0.45% NP-40 (IGEPAL)

0.45% Tween-20

0.01% Gelatin

Proteinase K:

20 mg/ml

10X PCR Buffer:

100 mM Tris, 500 mM KCl, 15 mM MgCl2 pH 8.3

dNTP mix:

25 mM/each


5-10 uM

Taq Polymerase:

approx 5U/ul


  1. Add proteinase K to lysis buffer (95 ul lysis buffer + 5 ul 20mg/ml proteinase K)
  2. Place 3 ul of lysis buffer in top of 200 ul PCR tube
  3. Pick single worm into lysis buffer
  4. Spin down to bottom of tube by spinning in microfuge 15 seconds @ 14,000 rpm
  5. Freeze tube @ -80 at least 1 hour
  6. Lysis of worm and release of genomic DNA
    • heat tube to 65 degrees for 60-90 minutes
    • inactivate proteinase K by heating to 95 degrees for 15 minutes
  7. Perform PCR
    • add 27 ul of PCR "master mix" to each tube
    • master mix: 1X PCR Buffer, 0.5 uM primers, 0.2 mM/each dNTPs, Taq polymerase
    • run PCR reaction for 30-35 cycles



compiled by Chad Rappleye

Aroian Lab Protocols