April+2012

=April 2012=

Preparation for agarose/arabinose plates
This month, I began by making some plates from scratch for the GFP lab that is going to be done by some high schools. On the plates, students will grow multiple colonies in which some will contain the GFP gene and others will not. I began using the protocol below: 1. Grab 2 liter flask and add 1L of pure water 2. Weigh out 40g/L of LB on weighing boat and add to flask 3. heat up LB in water in the microwave and mix the solution so that the LB dissolves 4. While LB solution cools, weigh and add 50mg of Ampicilian to 5mL of water to dilute 200x stock of 10mh/1mL ampicilian 5. Weigh and add 2g of arabinose to 10mL of water to dilute 100x stock of 200mg/mL arabinose L(+) 6. add both ampicilian and arabinose to LB solution to give ampicilian resistance to the plate
 * ~ Reagent ||~ Amount ||
 * Water || 1L ||
 * LB || 40g ||
 * Ampicilian || 50mg/5mL ||
 * Arabinose || 2g/10mL ||

Chemistry of Arabinose
What is the difference between L(+) and D(-) Arabinose? - The L(+) Arabinose will bind with the LB we are using but the D(-) will not because of their different chemical structures. What are chiral molecules? - They are a type of molecule that has a non-superposable mirror image, characterized as chirality ^[|chirality with hands] What are enantiomers? - They are steroisomers, those that differ only in spatial orientation, that are non-superposable mirror images of each.

Draw the chemical structure of L Arabinose and D Arabinose

Sectioning
This month I also sectioned a few more eye-cups that were injected with different inhibitors. I sectioned 1A and 2A obtaining about 10 slides of optic nerve for each to be sequenced for observation of proliferation due to the inhibitor. We received the results back and this sectioning was successful.

Zebrafish Eye


Above is a picture of a Zebrafish eye, visible is the lens, and three layers of the eye itself. The first layer is the ganglion and then Muller glia and then Photoreceptor cells. When sectioning we look for the optic nerve which runs through the center of the eye which is made up of ganglion cells that intertwine to make a thick rope like structure which is the optic nerve. The Zebrafish eye in its structure is extremely similar to the human eye and is therefore why we are sectioning and experimenting on them.