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Things we did today

Wetlab work

Margaret, Krystle, Norman

Today we made DH5-alpha competent cells. The first OD reading was between 0.3-0.4 for all three flasks. The OD after the last incubation with CMBD buffer was 1.1, 1.29, 0.8977, 1.079, and 0.942. (both of these OD readings are within the range we want.)

Margaret

oriT constuct from pSB1A2- for construction with J33207 for verification test
rep (pRL1383a)- to ligate to plac, B0030, and pSB1A3 and make a number of construct that can be sequenced with my new primers
omega interposon contruct from pSB1A2- to verify and sequence
oriV(pRL1383a)- to ligate to pSB1A3 and sequence next week.
(+) control pSB1A3 with VF2&VR
(-) control water with VF2&VR

Margaret

a= aadA
p=pRL1383a
b=from BB registry
first #= colony from aada+pSB1A3 ligation (sequence verified)
second#= colony from plac,B0034 ligation to the sequence verified aadA+pSB1A3 ligation

name of person/people who performed the task

Summary of task and what was done. Link to experiment for detailed notes if necessary.
e.g. read through papers, worked on proposal, etc.

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson

[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]

construct	Re-enzymes used	next step
J33207	SP	dephosphorylate
ap36	E, S	gel purify
ap32	E, S	gel purify
ap61	E, S	gel purify
ab95	E, S	gel purify
ab92	E, S	gel purify
ab96	E, S	gel purify
ab94	E, S	gel purify
ab98	E, S	gel purify
ab93	E, S	gel purify
base vector(I15020)	N	dephosphorylate
oriT(BB)	XP	ligate directly
oriT(my construct)PCR product 9/13	XP	ligate directly
TT(B0016)	XP	ligate directly
pSB1A3	EP	ligate to self(use as control)