Home›Genetics / Genomics›Extraction: High Molecular Weight Genomic DNA From Soils & Sediments l Protocol Preview
Steps
1Prepare denaturing buffer with beta mercaptoethanol00:56
2Assemble and chill mortar and pestle01:20
3Add soil sample and denaturing buffer to mortar01:30
4Cover sample with liquid nitrogen01:43
5Grind sample until powdery and homogeneous01:48
6Repeat grinding with additional liquid nitrogen01:56
Genetics / GenomicsJoVE (YouTube)
Extraction: High Molecular Weight Genomic DNA From Soils & Sediments l Protocol Preview
Protocol
Difficulty
intermediate
Steps
1
Prepare denaturing buffer with beta mercaptoethanol
Complete the denaturing buffer by adding beta mercaptoethanol to a concentration of 0.5 percent. Prepare the solution fresh or use buffer less than one week old kept at 4 degrees Celsius.
▶ 00:56
2
Assemble and chill mortar and pestle
Obtain a dewar of liquid nitrogen and prepare an autoclaved, pre-chilled mortar, pestle, and spatula for use in grinding the soil sample.
▶ 01:20
3
Add soil sample and denaturing buffer to mortar
Place a two-gram frozen soil sample in the pre-chilled mortar and add one milliliter of the denaturing solution.
▶ 01:30
4
Cover sample with liquid nitrogen
Add liquid nitrogen to the mortar to fully cover the soil sample.
▶ 01:43
5
Grind sample until powdery and homogeneous
Use the pre-chilled pestle to grind the sample until the soil particles appear powdery and homogeneous and the sample begins to melt.
▶ 01:48
6
Repeat grinding with additional liquid nitrogen
Add more liquid nitrogen to the mortar and repeat the grinding step to ensure complete homogenization.