Difference between revisions of "Team:Missouri Rolla"

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Revision as of 12:41, 17 October 2016

Defending North American bats from the emerging White-Nose epidemic

Here in Missouri and across the United States, bats play a major role in the ecosystem and economy. According to the USGS, bats are the primary consumers of insects in temperate regions. The insect suppression service that the bats provide saves the nation’s farmers between four and fifty billion dollars a year in lost crops and pesticide costs. Bats then produce waste that becomes the primary input of nutrients to Missouri’s over 7000 caves and allows the caves to support diverse and unique life.

Unfortunately, in 2007 a disease that is now called White Nose Syndrome (WNS) appeared in the bat populations of the northeastern United States. It is caused by a white fungus, Pseudogymnoascus destructans. The fungus grows on the bats while they hibernate and causes skin lesions leading to inflammation. This inflammation and irritation wakes the bats from their hibernation, causing them to burn precious fat reserves while it is still winter. The bats then starve or die from shock before spring arrives. The fungus has spread rapidly across the eastern United States and has left behind it high mortality. Generally, bat populations have declined by 80% with some populations totally lost.

[WHY NO ANTIFUNGALS, OCIMENE]

[LEUPEPTIN DESCRIPTION AND LINK TO PROJECT PAGE]

16 October 2016

Kira Buckowing

Start: 12:00pm

Chemical Transformations

Purpose: To check on the most recent work and get the wanted ocimene synthase in a chlor resistance backbone.

Protocol:

HiFi assemblies were set up as per the NEB protocol and incubated in ther thermocylcler for 15 minutes at 50 C.
10/16 HiFi 1
9 μL 10/14 D1
1 μL 10/14 D4
10 μL HiFi Master Mix
10/16 HiFi 2
9 μL 10/14 D2
1 μL 10/14 D4
10 μL HiFi Master Mix
14 broth Cultures were set up from colony growth on the 10/15 chemical transformation plates. Chemical transformations were performed using the chemically competent cells from NEB and adding either 2 μL or 6 μL of the appropriate HiFi assembly product, noted in the products section below.

Stop: 3:30pm

Products:

 
Label Source Description
10/16 CT1 100 μL 10/16 HiFi 1 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 2 μL added
10/16 CT1 250 μL 10/16 HiFi 1 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 2 μL added
10/16 CT2 100 μL 10/16 HiFi 2 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 2 μL added
10/16 CT2 250 μL 10/16 HiFi 2 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 2 μL added
10/16 CT3 100 μL 10/16 HiFi 1 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 6 μL added
10/16 CT3 250 μL 10/16 HiFi 1 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 6 μL added
10/16 CT4 100 μL 10/16 HiFi 2 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 6 μL added
10/16 CT4 250 μL 10/16 HiFi 2 Chemically transformed plasmid hopefully containing ocimene synthase in the pSB1C3 iGEM shipping backbone, 6 μL added
10/16 HiFi 1 10/14 D1 and D4 Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 HiFi 2 10/14 D2 and D4 Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC1 10/15 CT2 100 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC2 10/15 CT2 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC3 10/15 CT2 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC4 10/15 CT2 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC5 10/15 CT2 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC6 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC7 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC8 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC9 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC10 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC11 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC12 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC13 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone
10/16 BC14 10/15 CT4 250 μL Hopefully the assembled product of ocimene in the iGEM shipping backbone

Next: Minipreps of any growth in the broth cultures and checking the plates to see if anything grows.

Accordion test 2

Test 2