JNC44

On the creation of Biological SCP DNA Samples.

Over the years, I have been asked how I myself create and extract different DNA samples, and I will now share this with you all. It is in fact true that with even basic scientific skills you may create these samples, as long as you have the proper equipment and materials.

A Junior researcher has no less responsibility than a Senior, remember to always: follow protocol, take safety measures, and most importantly, secure, contain, protect.

On The Creation Of Biological Samples:

Homogenization

1 g of the biological material should be taken and cut into pieces, then ground using a porcelain mortar and pestle in 3 ml of lysis buffer containing 900 μl of 10% SDS. The emulsion should then be transferred to micro-centrifuge tubes and 100 μg proteinase K added per ml of emulsion solution, and incubated for 1 h at 50 °C.

Phase separation

350 μl of neutral saturated salt solution (NaCl) per ml should be added to the previous emulsion, the micro-centrifuge tube should be capped and shaken gently by the foundation's MX-42 (Easily acquired by filling in a request form to your Project Manager) for 15 s, and then incubated at room temperature for 10 min. The micro-centrifuge tube will then be centrifuged at 590 × g for 15 min at room temperature with DNA remaining exclusively in the aqueous phase

DNA precipitation

The resulting aqueous phase should then be transferred into another micro-centrifuge tube, and mixed with two volumes of room temperature absolute ethyl alcohol. Then the micro-centrifuge tube should be inverted several times for 10 s.

DNA wash

The supernatant should be removed; the DNA pellet should be washed once with 75% ethanol, and the DNA should be precipitated out of the solution by centrifugation at 9500 × g for 5 min.

On obtaining highest quality DNA:

The treatment of DNA with RNase should be done in a Tris buffer at the end of the extraction protocol. The steps can be repeated as before according to the protocol to obtain DNA with highest quality. The DNA can be precipitated and washed with 70% ethanol, and then the pellet can be dissolved in Tris-EDTA (TE) for DNA protection from degradation by metal dependent nucleases during storage.