ITS2_Box1anndBox4_PCR2_1

PCR 2 adding index to samples from Box 1 and 4 amplified for ITS2 marker

All steps following detailed instructions from Prada Lab PCR2 Protocol

  1. Move samples from strip tubes to 96 well replicate

  2. Standardize volumes with molecular Water Volume Plate

  3. 0.8X bead cleanup

  4. Add Index Volume Plate

  5. PCR 2

Thermocycler Settings:

Cycles Time Temp
1 3 min 98°
8 30 sec 98°
8 30 sec 55°
8 30 sec 72°
1 5 min 72°
1 ∞ min 4°
  1. 0.8X bead cleanup

  2. 1% Agarose gel to check for amplification Gel Image Gel Image

  3. Plate reader to determine DNA concentration
Written on July 7, 2022