Team:University of Ottawa/16 July 2008

From 2008.igem.org

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
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/16_July_2008"