IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week4/Chemical and Light: Difference between revisions
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| 45XP+38SP||AMP||4||DID NOT FLUORESCE- not miniprepped | | 45XP+38SP||AMP||4||DID NOT FLUORESCE- not miniprepped | ||
|- | |- | ||
| 51XP+38SP||CARB||5||2 picked colonies, neither grew- repicked 7/14 | | 51XP+38SP||CARB||5||2 picked colonies, neither grew- repicked 7/14, these did not grow either | ||
|- | |- | ||
| 54ES+59EX||KAN||3||DID NOT FLUORESCE- not miniprepped | | 54ES+59EX||KAN||3||DID NOT FLUORESCE- not miniprepped | ||
Revision as of 06:54, 15 July 2008
Primers
HO+pcyA, P3 vector w/o GFP
Primers were designed to PCR out the heme oxygenase and pcyA coding regions from the UT Austin plasmids. Due to the RE sites in these nonBB plasmids, their ends are given ApaLI and KpnI sites. The same is done for the P3 vector (the primers are such that we can PCR out the backbone, along with the RBS and terminator. The "P3-GFP" primers should also work for P1. However, I'm a little concerned, b/c the annotation of the P1 and P3 vectors disagrees with the sequence given for the RBS and terminator subparts.
P38, P39
5' phospho oligos were made to anneal into what is effectively P38 and P39 digested with ES.
Ligations
Results from 7/12 ligations (transformed into E1)
| DNA | Selection used on plate | # colonies | Notes |
| 17XP+38SP | AMP | 2 AFTER 48HRS | Plate put in 4°C, not picked |
| 63+59 FROM 7/9 LIGATION | KAN | 1 AFTER 48HRS | Plate put in 4°C, not picked |
| CDF XP (7/4)+P1XP | KAN | 1 AFTER 48HRS | Plate put in 4°C, not picked |
| 45XP(7/11 45)+39SP | AMP | 0 | |
| 45XP(7/11 45)+38SP | AMP | 0 | |
| 77XP+38SP | AMP | 0 | |
| 52XP+39SP | AMP | 0 | |
| 77XP+39SP | AMP | 0 | |
| 89XP+38SP | AMP | 0 | |
| 45XP+39SP | AMP | 0 | |
| 89XP+39SP | AMP | 0 | |
| 51XP+39SP | AMP | 0 | |
| 17XP+39SP | AMP | 0 | |
| P88 | KAN | 0 | |
| 63+58 FROM 7/9 LIGATION | KAN | 0 | |
| 58EX+63ES | KAN | 0 | |
| 52XP+38SP | AMP | 0 | |
| 89ES+45EX | CARB | TMTC | The 45 EX used was not purified, so the miniprep culture was of religated P45 and did not glow. However, we picked fluorescent colonies from the plate and restreaked/set up liquid cultures of them. |
| CDF XP(7/4)+P1XP(7/1) | KAN | 68 | miniprepped |
| 17ES+45EX | CARB | TMTC | The 45 EX used was not purified, so the miniprep cultures were of religated P45 and did not glow. However, we picked fluorescent colonies from the plate and restreaked/set up liquid cultures of them. |
| 45XP+38SP | AMP | 4 | DID NOT FLUORESCE- not miniprepped |
| 51XP+38SP | CARB | 5 | 2 picked colonies, neither grew- repicked 7/14, these did not grow either |
| 54ES+59EX | KAN | 3 | DID NOT FLUORESCE- not miniprepped |
| CDF XP(7/12)+P1XP(7/1) | KAN | TMTC | miniprepped |
| CDF XP(7/12)+P1 | KAN | 232 | miniprepped |
| no DNA | AMP | 0 | |
| no DNA | KAN | 0 | |
| no DNA | CARB | 6 big + 10 small | |
| no cells | KAN | 0 | |
| 1μL pUC19 | CARB | 96 | did grow in LB AMP medium (+ control) |
| 1μL pUC19 | AMP | 64 |
