IGEM:MIT/2006/Notebook/2007-1-11: Difference between revisions

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3. Write UROP Proposal- DONE
3. Write UROP Proposal- DONE


4. Perform colony PCR on many colonies from the J45120 and J45800 transformations- DONE and make LCs
4. Perform colony PCR on many colonies from the J45120 and J45800 transformations- DONE and make LCs- DONE


5. Track down Indole-Knockout cells
5. Track down Indole-Knockout cells- STILL LOOKING


6. Check for sequencing results
6. Check for sequencing results- DONE


7. Make a checklist for each part
7. Make a checklist for each part- DONE


8. Run gels from the colony PCR of J45120 and J45800 transformants
8. Run gels from the colony PCR of J45120 and J45800 transformants - ''J45120: most colonies were approx 1600 kB on the gel, which is correct. will read the gel for J45800 tomorrow''


'''Working List of Parts'''


Part              Description                                      Antiobiotic Test      Sequencing    Works
'''SEQUENCING RESULTS'''
 
*J45700 only had J45320 in it, but no J45120
J45120   Wintergreen Generating Device      Unsuccessful/retransformed and colony PCRed   
*J45320 looked correct
 
*J45180 was just the osmY + inverter, except no R0040 + WGD
J45180   Phase-Sensitive Wintergreen Generating Device              Successful            Pending
*J45200 looked correct, but the internal forward primer didn't work so we didn't get sequencing data for approx 150 base pairs in the middle. we'll do a smell test to make sure it's correct
 
*J45120 had an AAT insertion between 1197 and 1198, which should be ok. we'll do a smell test for that too.
J45200   Banana Generating Device                                  Successful            Pending
 
J45250  Phase-Sensitive Banana Generating Device                  Successful            Pending
 
J45320  Salicylic Acid Generating Device                          Successful            Pending
 
J45400  Isoamyl Alcohol Generating Device                          Successful            Pending
 
J45700  Autonomous Wintergreen Generating Device                  Successful            Pending
 
J45800  Autonomous Phase-Sensitive Wintergreen Generating Device  Transformed
 
J45999  Indole-Knockout Cells                                      Looking for it
 
pBANANA  Autonomous Banana Generating Device                        Unsuccessful

Latest revision as of 09:19, 12 January 2007

Plating Results from Yesterday

1. Jason's parts finally grew up. The control, B0015, worked only on the Kan plate as expected (it is in the 1AK3 plasmid). Mach I interestingly grew up on Kan and Tet but not Chl. pBANANA #10 and #12 both grew up on Kan and Chl but not on Tet.

2. J45180 only grew up on AMP/KAN as expected, confirming that the cell is unlikely to have plasmids of different resistances in it.

3. Interestingly, J45400, which was streaked out by Meagan on an AMP/KAN plate did not grow on AMP/KAN, AMP/TET, or AMP/CHL. I will repeat this experiment today.

4. Many colonies were seen from the transformations of J45120, the two J45800 hopefuls, and pUC18.

Plan Today

1. Miniprep, glycerol, and sequence J45400- DONE

2. Repeat streaking of J45400 onto AMP/KAN, AMP/TET, and AMP/CHL- DONE

3. Write UROP Proposal- DONE

4. Perform colony PCR on many colonies from the J45120 and J45800 transformations- DONE and make LCs- DONE

5. Track down Indole-Knockout cells- STILL LOOKING

6. Check for sequencing results- DONE

7. Make a checklist for each part- DONE

8. Run gels from the colony PCR of J45120 and J45800 transformants - J45120: most colonies were approx 1600 kB on the gel, which is correct. will read the gel for J45800 tomorrow


SEQUENCING RESULTS

  • J45700 only had J45320 in it, but no J45120
  • J45320 looked correct
  • J45180 was just the osmY + inverter, except no R0040 + WGD
  • J45200 looked correct, but the internal forward primer didn't work so we didn't get sequencing data for approx 150 base pairs in the middle. we'll do a smell test to make sure it's correct
  • J45120 had an AAT insertion between 1197 and 1198, which should be ok. we'll do a smell test for that too.
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