Joshua Harrison

Plant samples from 2 replicates of 50 individuals were planted in 19 common gardens across the USA. This will explore the interaction of plant species, microbiome, and climate. We will explore some cheaper extraction and PCR methods for library prep.

Sample sheet can be found here.

Extractions

• Try the cheaper Machery-Nagel kits out - making 10 duplicates to try MN membrane filter kit

Organization

• Naming Convention: 2021-UWEC-411-2

  • Year-Garden-Species-Block

    • Blocks should be 1 or 2, any other number is likely row

      • Add “R” before last number for all samples in a bag with numbers higher than 2: 2021-UWEC-321-R3

• Deal with each outer bag together to check for errors

  • Each Garden should have 2 blocks

  • Each block should have one of each species ID

  • Each Block is arranged 5x10

Washing

1. Load sample in tube

2. Fill half way with Ultra Pure Water

3. Shake in Mixer Mill (or Vortex?)

4. Dump out Liquid

5. Continue from Step 2 once

6. Save both washes separately for ?half? of the samples

Library Prep

Explore Extraction Free PCR from lysates

Taq Candidates:

• Hemo KlenTaq

• Phire Plant Direct

• FastAmp® Plant Direct PCR Kit