Thursday, April 30, 2015

Week 15. Semester 2. Happy hour!

This week I retrieved my fungal cultures from the agitator and attempted to isolate the exoenzymes for use in DNA extraction later. The broth was centrifuged for 30 min at 5000 rpm and then filtered with a .25 micrometer paper in a vaccum. Only two of the broths filtered successfully and the other two plugged the smaller .25micro filter and a larger .45 micrometer filter as well, so they have not been filtered.
Additionally, another DNA extraction protocol was attempted and gave very promising results! In a nut shell, the protocol uses a 2.5% SDS lysis buffer and a one step 1:1 Phenol-Chloroform extraction (Lopez et all. 2013). This week has also been spent preparing my poster for the Estrella Mountain Student Conference.



 Vaccum Filtration for centrifuged media.
Top lanes are from new protocol ad bottom from previously used one. 


Lopez M, Gonzalez-Mendoza D, Grimaldo-Juarez O. 2013. Fast protocol for extraction of DNA from Prosopis spp leaves (plant adapted to arid environment) without liquid nitrogen. Genetics and molecular research [Online];[cited 03/04/2015]. 12.3 (4090-4094)Available from: http://www.funpecrp.com.br/gmr/year2013/vol12-3/pdf/gmr2488.pdf
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2 comments:

  1. AnonymousMay 12, 2015 at 1:17 PM

    Congrats on your poster win! I was wondering how you plan on preserving your exoenzyme?

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  2. ZGrijalvaSeptember 25, 2015 at 9:21 AM

    Thank you Cori!!! I barely found this comment hehe
    at the moment I haven't resumed the experiment nor successfully or fully isolated the enzyme. There are some special filters I need for the centrifuge. If I do continue with the experiment I will definitely need some help with preserving the enzymes!! Thank you for all your help in the Lab Cori!

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