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Team:Hawaii/Notebook/2008-10-23
From 2008.igem.org
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(→Large scale plasmid prep) |
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:* No change from yesterday | :* No change from yesterday | ||
:* Colony PCR of 6 colonies from sp<sub>40, 50, 75</sub> and 8 colonies from sp<sub>100</sub> plates | :* Colony PCR of 6 colonies from sp<sub>40, 50, 75</sub> and 8 colonies from sp<sub>100</sub> plates | ||
| + | ::* sp<sub>75,100</sub> colonies correct | ||
:* Restreaked sp<sub>2.5</sub> blue colony, sp<sub>100</sub> #1 & 2 on LB+amp<sub>100</sub>, LB, LB+sp<sub>100</sub> + X-gal | :* Restreaked sp<sub>2.5</sub> blue colony, sp<sub>100</sub> #1 & 2 on LB+amp<sub>100</sub>, LB, LB+sp<sub>100</sub> + X-gal | ||
| Line 16: | Line 17: | ||
:*From 9/28 sp<sub>2.5</sub> plate: #1 | :*From 9/28 sp<sub>2.5</sub> plate: #1 | ||
::* Gel showed no DNA in lanes migrated. Huh? Will try one more time to plasmid prep tomorrow. | ::* Gel showed no DNA in lanes migrated. Huh? Will try one more time to plasmid prep tomorrow. | ||
| + | |||
| + | ===Inoculated for plasmid prep=== | ||
| + | :<strong>Grace</strong> | ||
| + | :* In 10 ml TB+amp<sub>100</sub>: | ||
| + | ::* nir | ||
| + | ::* nir+rbs | ||
| + | ::* slr1 | ||
| + | ::* pilA | ||
| + | ::* GFPf | ||
| + | ::* slr1+GFPf | ||
| + | :* In 50 ml TB+sp<sub>100</sub>: | ||
| + | ::* BB1 (sp<sub>100</sub> plate) | ||
| + | :* Incubated at 37C for 18 hours with shaking at 250 rpm | ||
===Verification of transformants=== | ===Verification of transformants=== | ||
| Line 31: | Line 45: | ||
|align=center|0 | |align=center|0 | ||
|- | |- | ||
| - | |align=center|BB-1 | + | |align=center|BB-1 (plasmid from 9/28 plate) |
|align=center|4 | |align=center|4 | ||
|- | |- | ||
| Line 45: | Line 59: | ||
:* EcoRI/PstI: nir+rbs | :* EcoRI/PstI: nir+rbs | ||
:* XbaI/SpeI: slr1+GFPf | :* XbaI/SpeI: slr1+GFPf | ||
| + | |||
| + | ===Plasmid Prep=== | ||
| + | :<strong>Krystle</strong> | ||
| + | :*Plasmid prep of | ||
| + | :**lac+rbs+slr+gfpf #1,3,4 | ||
| + | :**nir+rbs+slr+gfpf #2,3 (from last week) | ||
= Discussion = | = Discussion = | ||
Latest revision as of 01:07, 30 October 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Antibiotic test
- Grace
- No change from yesterday
- Colony PCR of 6 colonies from sp40, 50, 75 and 8 colonies from sp100 plates
- sp75,100 colonies correct
- Restreaked sp2.5 blue colony, sp100 #1 & 2 on LB+amp100, LB, LB+sp100 + X-gal
Large scale plasmid prep
- Grace
- From sp100 plate: #1, 2
- From 9/28 sp2.5 plate: #1
- Gel showed no DNA in lanes migrated. Huh? Will try one more time to plasmid prep tomorrow.
Inoculated for plasmid prep
- Grace
- In 10 ml TB+amp100:
- nir
- nir+rbs
- slr1
- pilA
- GFPf
- slr1+GFPf
- In 50 ml TB+sp100:
- BB1 (sp100 plate)
- Incubated at 37C for 18 hours with shaking at 250 rpm
Verification of transformants
- Grace
| Plasmid | Colonies |
|---|---|
| J04430 | 0 |
| (-) on amp100 | 0 |
| BB-1 (plasmid from 9/28 plate) | 4 |
| (-) on sp100 | 0 |
- Colony PCR of 4 BB-1 transformants
- None were correct
RE digest
- Grace
- EcoRI/PstI: nir+rbs
- XbaI/SpeI: slr1+GFPf
Plasmid Prep
- Krystle
- Plasmid prep of
- lac+rbs+slr+gfpf #1,3,4
- nir+rbs+slr+gfpf #2,3 (from last week)
- Plasmid prep of
Discussion
Quote of the Day
History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
