15-16 April 2017 workshop

We had 11 kids from 8 to 15 years old joining the 2 day workshop. In the workshop, they collected 33 plant samples in total, and then using some basic molecular biology lab skills including DNA extraction, PCR and gel electrophoresis to generate the barcode of these plant samples.

I. Collect samples and document:

We use iNaturalist to document our collections. Photos, geolocation and notes are added to the DIYBIOHK Barcode Hong Kong Project.

Note: 15/04 #1-33

Number of samples: 33

II. Isolate DNA from plant samples

We used the DNeasy Plant kit from qiagen. Each students worked on 3 samples.

III. Amplify DNA barcode by PCR

4ul DNA template was added to each pcr tube

amplify rbcL gene

Reaction mixture:

PCR profile:

Since the limited place in the PCR machine, we only pcr 20 samples at the workshop. So we still need to do 2nd bench of PCR to finish all 33 samples.

IV. Analyze PCR products by Gel electrophoresis

Gel 1

Gel 2

Fig. 1 Gel photo of PCR products. 7ul pcr products was added in each well. The DNA barcode (rbcL) gene is ~600-700bp.

The DNA bands are too thick, it is recommended to load less volume (5ul) sample to the well. The DNA ladder is blurry, may be the gel electrophoresis was running too fast or the DNA ladder was degraded.

Next step:

- Send the successful pcr products (barcode) to do DNA sequencing.

- Develop a more economic DNA extraction method, with reference to the Barcoding 101 protocol.