Wednesday, February 17, 2016

bio125 20160217Wed RE of PCR fragments

Before class:
Set 4 water bath/blocks: 60C,  37C, 55C heat block

Water is on their stations. Enzymes are in small -20 in the green ice block on top shelf. Buffers are in 2 boxes on the top shelf labeled buffers for RE enzyme and Restriction enzyme buffer. Each student can get a tube of the buffer that they need. After assembling MMix, the instructor should dispense the enzyme into their master mix for digestion. Water bath are set to 37 and 60. For 55 degree they should use heating block which is turned on and set.

Student samples are in the freezer by the tissue culture hood in the front with section labeled on the box

NEB reformulated their buffers. 
Buffer 2.1 = old buffer 2 + BSA

Section 3:
9:10, go over the lab.  I did not put a tutorial video in the right place, and most group did not know which RE to use. I used a student video in class to let the student work on it.

by 10am, most students worked out the solutions.

Section 4
No groups submitted a preclass report that indicated RE needed.
Go over student video on RE identification
Showed student video from morning section.

by 1:41pm, three groups correctly found out RE needed.
by 2:12pm,  all groups knew their RE.

2:20pm, all groups start to set up reactions. One group was absent during last lab and did not do PCR. They were asked to finished APE 9 assignment.

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