Silage protocols
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Introduction
These protocols are use for the analysis of any forage material, but especially silage.
pH Measurement
- Weigh out 5g of silage.
- Add this material to 45ml of nanopure water
- Put in the shaker for 30 mins.
- Measure pH by submerging the probe in the liquid.
Plate Count
- 1. Add 25g of silage to 25ml of TE Buffer in a small blender.
- 2. Blend 2X for 10 seconds (each time).
- 3. Weigh 2g of this mix and add to 8ml DI water (This the first 1:10 dilution of your original sample).
- 4. Vortex to mix.
- 5. Perform five more 1:10 serial dilutions by adding 100ul of the previous dilution to 900ul water.
- You will use the last three of these dilutions for plating (1:104,1:105, 1:106).
- 6. Plate 100ul of each of these three final dilutions in triplicate.
- This means you'll use a total of nine plates.
- 7. Calculate the total count per 1g of silage.
- For 1:104 dilution multiply the plate count by 105 to get cfu/g silage.
- For 1:105 dilution multiply by 106.
- For 1:106 dilution multiply by 107.
- If a plate has more than ~300 colonies,then forget about that plate.
- 8. Average results.
DNA Extraction for QPCR
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