IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/10/23

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July 23rd, 2009

Qubit Fluorometer Test of PCR Products

  1. DNA Concentration x 200

1. 0.295 ug/ml -> 59 ug, ml

2. 0.345 ug/ml -> 69 ug/ml

3. 0.396 ug/ml -> 79 ug/ml

4. 0.301 ug/ml -> 60 ug/ml

5. 0.417 ug/ml -> 83.4 ug/ml

used standard protocol . working buffer = 199 ul, DNA = 1 ul

End : 12:30

Agarose Gel to Confirm Successful PCR x 1.2% agarose

- Controls (7+8) both did not work! - DNA fragments all present in proper positions (2.3 kb) - DNA ladder a bit lacking in concentration

End 1:30

Transformation of Longtine PCR Products

Followed usal procedures, but left overnight in rollers of 500 ul each culture in order to boost growth after heat shock. Plated on -His plates on 7.24 in the morning. After heatshock, do not suspend in SOS, but rather resuspend in 500 ml YPD and rotate overnight. May improve cell efficiency.