Team:University of Chicago/Notebook/Norayucel

From 2008.igem.org

(Difference between revisions)
(→July 18, 2008)
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10. Streaked 20microliters onto Amp. Plates<br>
10. Streaked 20microliters onto Amp. Plates<br>
11. Dan will come in Saturday morning to pick up the plates and count colonies.<br>
11. Dan will come in Saturday morning to pick up the plates and count colonies.<br>
 +
 +
==July 25, 2008==
 +
#Remade TOP10 competent cells--will try to make more competent
 +
#Inoculated 500mL of SOB at ~10am with 1mL each of TOP10 starter culture
 +
#*Two 2L flasks with 250mL each
 +
#At 5:30 OD was .28 and .29 for each flask respectively
 +
#Final OD after following competent cell protocol was 1.04 (between 1-1.5 as required)
 +
#Put into -80C at 7:30pm.
 +
 +
==July 28, 2008==
 +
'''Transformation with pGreen'''
 +
#10pg/microliter on LB Amp
 +
#1 ng/microliter on LB Amp
 +
#NO plasmid on LB Amp
 +
#NO plasmid on plain LB
 +
#Put into 37C incubator ~1:30pm<br><br>
 +
'''Plates'''
 +
#Running out of plates
 +
# 500mL of LB and LB Amp agar
 +
#*100mg/mL stock of Ampicillan
 +
#* After cooling for ~45 minutes at room temp, added .5mL
 +
#Poured around 4:30 pm. Turned over at 5:30<br>
 +
 +
Checked plates before I left at ~6pm. No growth (as expected). Will look again in the morning
 +
 +
==July 29, 2008==
 +
#Checked plates
 +
# 10 pg/microliter plate had ~20 colonies. Competency still off by factor of 10 :(
 +
#1ng/microliter plate was a big streaky mess (too many to count)
 +
#NO plasmid on LB+Amp also big streaky mess. Possibly mislabeled plate??
 +
#NO plasmid +LB big streaky mess (as expected)

Revision as of 16:56, 29 July 2008

Contents

July 18, 2008

1. Diluted 1ng/microliter pGreen plasmid to 10pg/microliter with 100X dilution
2. 1microliter of plasmid to 50 microliters competent cells
3. Will grow up 50microliters of TOP10 untransformed as a control
4. Put on ice at 2:03
5. Took off ice at 2:34
6. Incubated for 1minute at 42C
7. Added 250microliters SOC media (prepared at 1:30)
8. Start incubated at 37C at 2:36
9. Take out at 3:40pm
10. Streaked 20microliters onto Amp. Plates
11. Dan will come in Saturday morning to pick up the plates and count colonies.

July 25, 2008

  1. Remade TOP10 competent cells--will try to make more competent
  2. Inoculated 500mL of SOB at ~10am with 1mL each of TOP10 starter culture
    • Two 2L flasks with 250mL each
  3. At 5:30 OD was .28 and .29 for each flask respectively
  4. Final OD after following competent cell protocol was 1.04 (between 1-1.5 as required)
  5. Put into -80C at 7:30pm.

July 28, 2008

Transformation with pGreen

  1. 10pg/microliter on LB Amp
  2. 1 ng/microliter on LB Amp
  3. NO plasmid on LB Amp
  4. NO plasmid on plain LB
  5. Put into 37C incubator ~1:30pm

Plates

  1. Running out of plates
  2. 500mL of LB and LB Amp agar
    • 100mg/mL stock of Ampicillan
    • After cooling for ~45 minutes at room temp, added .5mL
  3. Poured around 4:30 pm. Turned over at 5:30

Checked plates before I left at ~6pm. No growth (as expected). Will look again in the morning

July 29, 2008

  1. Checked plates
  2. 10 pg/microliter plate had ~20 colonies. Competency still off by factor of 10 :(
  3. 1ng/microliter plate was a big streaky mess (too many to count)
  4. NO plasmid on LB+Amp also big streaky mess. Possibly mislabeled plate??
  5. NO plasmid +LB big streaky mess (as expected)
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