An example of how some context would improve a protocol

protocol.md

Isohelix DNA Isolation Kit

• product page
• official instructions

Preparation

• Preheat the EB (Elution Buffer) buffer at 700C (100μl per sample)
• Set centrifuge to 13.4K rpm (12,000g), it is the only speed we will use

Collect DNA sample

• Swab inside of cheek for thirty seconds
• Place the swab head into a tube

Remove DNA from cells

• Add 500μl LYS lysis buffer, which breaks down the cells containing the DNA
• Vortex to cover the swab head
• Add 20μl Proteinase K solution, which digests (eats) contaminating proteins
• Mix immediately by vortexing
• Incubate at 600°C for 10 to 60 minutes

Unknown things for unknown reasons

• Add 750μl CB (Column Binding) buffer, which maybe helps the DNA stick to the filter or something.
• Vortex thoroughly (at least 30 seconds)
• Add 1.25ml ethanol to the sample, maybe this kills anything the lysis and Proteinase K missed?
• Vortex to mix

Remove DNA from solution

The solution is full of dead cells and proteins, it's basically cell and protein graveyard, kinda spooky, lets get our DNA out! We'll run it through a filter called a "column".

Anatomy of a column:

• A tube that is the postfilter basin
• A tube that is the prefilter basin, which fits inside the postfilter basin
• At the bottom of the prefilter basin, a faucet to drain it
• Within the faucet, a membrane that acts as the filter

We'll run our sample through it, 700μl at a time, catching the DNA on the membrane and let the ghostwater flow through.

• Repeat until the entire sample is processed:
  • Add 700μl of the sample to the prefilter basin, be careful not to touch the rim of the faucet.
  • Centrifuge for 1 minute
  • Discard the flow-through

Rinse the DNA twice and dry it off

• First rinse
  • Add 750μl solution WB
  • Centrifuge for 1 minute, so WB flows through
  • Discard the flow through
• Second rinse
  • Add 750μl solution WB
  • Centrifuge for 1 minute, so WB flows through
  • Discard the flow through
• Dry
  • Place the column in a clean collection tube (postfilter basin) since the current one is all wet
  • Centrifuge 3 minutes, to remove all traces of ethanol (presumably that's the WB?)

Elution: Remove the DNA from the membrane

In the next step, we "incubate", which is defined as "(especially in a laboratory) keep (eggs, cells, bacteria, embryos, etc.) at a suitable temperature so that they develop", but we only have DNA and EB, so are we incubating the EB? Is it al

• Place the column in a clean 1.5ml microcentrifuge tube
• Add 100μl of preheated EB (Elution Buffer) buffer to the center of the membrane
• Incubate the column for 3 minutes (are they using "incubate" to mean "heat", or is my DNA at risk of hatching, or is EB a bacteria or something?)
• Centrifuge for 1 minute to elute the DNA

Store the DNA

• Store the eluted DNA at –200C to preserve it

Xtreme_Kit_Instructions.pdf