Abel Thomas

2/7/19

Purpose: During this lab time, we started using metagenomics to get the DNA from the tree and the soil. We will sequence this DNA and also we will find the ciliates from the soil.

Materials:

Petri Dish

Eppendorf Tubes

Soil Sample

Dissecting Microscope

Compound Microscope

Silica beads

DNA extraction buffer

Pestle and Mortar

Heat Block

Falcon Tube

Ruler

Charcoal

Procedure:

1. We observed the leafs under the dissecting microscope to see if we could find anything that we couldn’t see from our first observation
2. Observe the characteristics of the leaf
3. Get falcon tube from the previous lab
4. Measure the soil, clay, and silt
5. Add 1 gram of the dry soil from the small plate and 1 gram of silica beads and grinded it with mortar and pestle for 5 minutes… Bassam’s soil was chosen because it had the most ciliates that were the best to see
6. Continue to grind with the mortar and pestle and add 10 mg of charcoal, after a minute add 2 mL of the DNA extraction buffer and keep grinding the mortar and pestle
7. Take pippete and put mixture from mortar and pestle into eppendorf tube
8. Put the eppendorf tube into the heat block for 10 minutes at 65 degrees Celsius
9. Spin the eppendorf tube in a vortex for 5 minutes
10. pippete supernatant from eppendorf tube without getting any soil

Data:

There was 1 cm of sand, .8 cm of silt and .3 cm of clay. which means there was 48% sand, 38% silt and 14% clay. this makes our soil to be sandy loam

The leaf was oblonceolate, an obtuse top, a rounded bottom, entire edges, odd pinnately compound

We determined the tree was a Texas Red Oak or Quercus Buckleyi

Storage: We put all our instruments in their appropriate spots and made sure to wash and clean our area and the instruments

Conclusion/Future Goals: We were able to move farther in the metagenomics process. We were able to extract the DNA. Our future goal is to sequence the DNA and help us idetnify the trees and the ciliates around the trees.