Silage protocols

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Introduction

These protocols are use for the analysis of any forage material, but especially silage.

pH Measurement

1. Weigh out 5g of silage.
2. Add this material to 45ml of nanopure water
3. Put in the shaker for 30 mins.
4. 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)

1. Plate 100ul of each of these three final dilutions in triplicate

• this means you'll use a total of nine plates.

1. 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.

1. Average results

References

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