Amplification of human genomic DNA

Last time we extracted our own genomic DNA. To determine whether the extracted DNA is of good quality, we tried to amplify a fragment of the human PDE1C gene by Polymerase Chain Reaction (PCR).

The two primers used were:

PDE1C_for: 5′-CTGGAACCAGTGCCACTATAA-3′; melting temperature of 54oC.

PDE1C_rev: 5′-CGACTTCAGCCATCTCTCTATC-3′; melting temperature of 55oC.

The conditions used for the PCR are accessible here.

We were expecting a 314 bp long fragment and the PCR products were run on an 1% agarose gel.

PCRgDNA_191114b

As expected, there was no band observed in the Negative Control since the kiwi does not contain the PDE1C gene which is always a good news. For our own genomic DNA, only the PC and M lanes produce the expected band. For the Y lane, we observed two bands at 100 bp which should correspond to some amplified DNA since the primers are only 21 bp long.

Since not all the extracted DNA worked as expected, we will try a new DNA extraction protocol.

Fingerers crossed we’ll get a hold of cheap method for extracting genomic DNA using kitchenware materials without the reliance on professional kits! This hopefully will give us the ability to test our own DNA for interesting genetic variants!

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