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Team:University of Ottawa/16 July 2008
From 2008.igem.org
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:<li>Control is LB + AMP only. | :<li>Control is LB + AMP only. | ||
:<li>Finished Innoculation at 3:oo pm | :<li>Finished Innoculation at 3:oo pm | ||
| + | '''Chris''' | ||
| + | :'''Gel Extraction of AtCRE''' | ||
| + | ::<li> prepared a 0.8% gel and ran the AtCRE sample for 40 minutes at 90 V | ||
| + | ::<li> expected bands appeared and the desired one was excised | ||
| + | ::<li> gel extraction was performed on the excised sample successfully | ||
| + | :'''Determining AtCRE Concentration''' | ||
| + | ::<li> measured the absorbancy of AtCRE, resulting in invalid data. It was determined that the blank was performed incorrectly, such that the machine was not zeroed properly. | ||
| + | ::<li> the absorbancy of AtCRE was remeasured using a new blank at a 1:10 dilution. The resulting absorbancy was 0.3113 at 260 nm, giving a concentration of about 150 ng/ul. | ||
| + | :'''Ligation of AtCRE''' | ||
| + | ::<li> AtCRE was ligated with 1 ul ligase, 1 ul ATP, 2 ul DNA template, 1 uL buffer and 5 ul water | ||
| + | ::<li> the sample was incubated at 16 C overnight | ||
Revision as of 20:04, 16 July 2008
Contents |
Today on the Lab
Tammy
- Innoculation of transformed X10 E.coli + pDR197::AtCKX2 in LB + AMP.
- For each dilution (PURE, 1:10, 1:100), 2 colonies were chosen to innoculate in 3 ml LM + AMP.
- Control is LB + AMP only.
- Finished Innoculation at 3:oo pm
Chris
- Gel Extraction of AtCRE
- prepared a 0.8% gel and ran the AtCRE sample for 40 minutes at 90 V
- expected bands appeared and the desired one was excised
- gel extraction was performed on the excised sample successfully
- Determining AtCRE Concentration
- measured the absorbancy of AtCRE, resulting in invalid data. It was determined that the blank was performed incorrectly, such that the machine was not zeroed properly.
- the absorbancy of AtCRE was remeasured using a new blank at a 1:10 dilution. The resulting absorbancy was 0.3113 at 260 nm, giving a concentration of about 150 ng/ul.
- Ligation of AtCRE
- AtCRE was ligated with 1 ul ligase, 1 ul ATP, 2 ul DNA template, 1 uL buffer and 5 ul water
- the sample was incubated at 16 C overnight
