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	===Labs===		===Labs===
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		+	Rebecca and Lukas went back into the labs today in order to try and transform the ''E. coli'' once more. Using the knowledge we had gained through research into why the previous transformation had been unsuccessful agar was used that had a reduced amount of chloramphenicol in order to try and grow more cells. Two controls were also set up in order to point us in the right direction with what may have gone wrong; the positive control used BRAF (BL21 pLysS) cells which contain vectors for chloramphenicol resistance; the negative control used the NEB 5-alpha cells used in the previous experiment; our aim was for the positive control to show whether the issue lies within the chloramphenicol resistance, or whether the problem lies within the competent cells themselves.
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		+	A sample of lysogeny broth (LB) as also inoculated with pre-made culture containing the BL21 pLysS cells in preparation for the next day's plasmid isolation focusing on chloramphenicol resistance.
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		+	Protocols for the experiments can be found [[Team:NRP-UEA-Norwich/Protocol#Transformation_2 | Here]]
	==Day 2==		==Day 2==

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Revision as of 11:51, 19 July 2012

Welcome to the NRP UEA iGEM 2012 Wiki Lab Book

Please choose the relevant link to access our diary of that week!

Week Zero | Week One | Week Two | Week Three | Week Four | Week Five | Week Six | Week Seven | Week Eight | Week Nine | Week Ten | Week Eleven | Lab Protocols | Experiments

Contents 1 Week 2 1.1 Day 1 1.1.1 Research 1.1.2 Labs 1.2 Day 2 1.3 Day 3 1.4 Day 4 1.5 Day 5

Week 2

DESCRIPTION OF WEEK TWO TO BE UPLOADED AT A LATER DATE

Day 1

Research

While two members of the team worked in the labs on the transformation of Escherichia coli the rest of the team continued with the research into previous BioBricks and those that we felt the characterisation of would be useful to our project. As one of our proposed ideas for the project is to produce a BioBrick that can sense nitric oxide and then go on to increase nitric oxide amounts to toxic levels if appropriate as a potential cancer therapeutic we decided to look into genes for nitric oxide synthase and nitrite reductase, both enzymes that result in the production of nitric oxide. We also looked into plasmids and genes that may be used in mammalian cells as well as bacterial in order to start thinking about where our project could go once the BioBrick is synthesised.

Labs

Rebecca and Lukas went back into the labs today in order to try and transform the E. coli once more. Using the knowledge we had gained through research into why the previous transformation had been unsuccessful agar was used that had a reduced amount of chloramphenicol in order to try and grow more cells. Two controls were also set up in order to point us in the right direction with what may have gone wrong; the positive control used BRAF (BL21 pLysS) cells which contain vectors for chloramphenicol resistance; the negative control used the NEB 5-alpha cells used in the previous experiment; our aim was for the positive control to show whether the issue lies within the chloramphenicol resistance, or whether the problem lies within the competent cells themselves.

A sample of lysogeny broth (LB) as also inoculated with pre-made culture containing the BL21 pLysS cells in preparation for the next day's plasmid isolation focusing on chloramphenicol resistance.

Protocols for the experiments can be found Here

Day 2

Day 3

Day 4

Day 5