Kicking off a new project this week with the brilliant Erik Roldan. Lets see if the student has surpassed the master running gels...
Previously our lab has seen a relationship between Wolbachia and the Liberibacter bacterium that causes citrus green disease. So we're going to use the FANA-ASO technology to disrupt a nutrient based interaction. There are a few genes being investigated to target. Erik did RT-PCR and then I used GoTaq Green for amplification. Which bring us to todays gel wars. We've got 4 gene products, which lucky winner will be sent off for sequencing?
Because of some solid bands in an old gel, Erik was expecting to see something in this top gel. However, the bottom gel should have been blank. So, I re-ran the gels with a higher protein concentration getting quite the same result. Nothing in the expected. So we pulled the original cell line DNA extraction and whattt?!
Lanes:1 DNA Ladder; 2-5 NTC; 6-9 Plant; 10,11 ACP; 112,13 Wolbachia.
Houston we have a Winner!!
Apparently it takes three times the DNA concentration to see these bands. (230ng/ul verses 70ng/ul)
New question: Why didn't he see the 2nd protein at all when he ran it yet it was the strong in my DNA?
- Same primers, same probes
This week there's going to be some troubleshooting then taking things to the boss. Which does she want to send for sequencing?
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