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Project:Plant species testing: Difference between revisions
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* Electrophoresed for 1 hr - started at 80V, settled at around 130V | * Electrophoresed for 1 hr - started at 80V, settled at around 130V | ||
* Visualised under UV light through orange filter - did not see any bands from the template, but ladder was visible, indicating PCR failed. | * Visualised under UV light through orange filter - did not see any bands from the template, but ladder was visible, indicating PCR failed. | ||
==== Repeat without PCR ==== | |||
Extraction: | |||
* 'Homogenize' (mush) approx 50 mg pea with a disposable pipette tube in 600ul of 5% chelex solution for 1 min | |||
* Vortex for 10s then boil for 5 mins, then vortex again for 10s. | |||
* Centrifuge for 1 min - recover supernatant and use as template | |||
Electrophoresis: | |||
* 100ml of agarose solution: 1 gram of agarose powder added to 100ml of 1% TBE solution | |||
* Microwaved the agarose, cooled to 50 degrees, added 5ul of ethidium bromide stock solution | |||
* Poured agarose into mould and waited to cool, then covered with 1% TBE. | |||
* Put ladder into far well, template into near well | |||
* Electrophoresed for 1 hr at 80V | |||
* Visualised under UV light through orange filter - saw a faint, blurry band, running ahead of the ladder, which we assumed to be the genomic DNA we were looking for. | |||
== 2nd go == | == 2nd go == | ||
