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Team:Hawaii/Notebook/2008-07-15
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:* Ligated 7 μl Biobrick segment with 0.5 μl pRL1383a and 7 μl GFP fusion with 2 μl BBa_C0012 vector in 20 μl ligation reactions | :* Ligated 7 μl Biobrick segment with 0.5 μl pRL1383a and 7 μl GFP fusion with 2 μl BBa_C0012 vector in 20 μl ligation reactions | ||
:* Transformed DH5α using 10 μl ligation reactions | :* Transformed DH5α using 10 μl ligation reactions | ||
| + | ::* Plated GFP/C0012 vector on LB + amp<sub>100</sub> | ||
| + | ::* Plated BB segment/pRL1383a on LB + sp<sub>100</sub> | ||
:* Counted colonies from yesterday's pnir, slr2016, pilA assembly transformation (see [[Team:Hawaii/Initial_Synth._Oligo_Assembly#Sixth_attempt|experiment]]) | :* Counted colonies from yesterday's pnir, slr2016, pilA assembly transformation (see [[Team:Hawaii/Initial_Synth._Oligo_Assembly#Sixth_attempt|experiment]]) | ||
:* Colony PCR'd colonies to verify presence of desired products | :* Colony PCR'd colonies to verify presence of desired products | ||
| Line 19: | Line 21: | ||
<strong>Margaret</strong> | <strong>Margaret</strong> | ||
| - | :*Transformation of DB3.1 competent cells pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-) no plasmid, because transformation from 7/14/08 yielded only 1 colony from pSB3K3, nothing from pSB1A2 & pSB1AK3. | + | :*Transformation of DB3.1 competent cells pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-) no plasmid, because transformation from 7/14/08 yielded only 1 colony from pSB3K3, nothing from pSB1A2 & pSB1AK3. |
| + | |||
| + | ===Plasmid prep=== | ||
| + | <strong>Krystle</strong> | ||
| + | |||
| + | :* Resuspended yesterday's plasmid prep in 100 μl TE and stored in -20C | ||
| + | :* Ran plasmid preps on gel to verify | ||
| + | :* Grew up pnir, slr2016, and pilA transformants in LB in preparation for plasmid prep tomorrow | ||
| + | |||
| + | ===Restriction Digest=== | ||
| + | <strong>Krystle</strong> | ||
| + | |||
| + | :* RE digested BBa_B0034, BBa_B0024, and BBa_E0040 with XbaI overnight | ||
= Discussion = | = Discussion = | ||
Latest revision as of 18:21, 16 July 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Oligonucleotide experiment, continued
- Grace
- Ran overnight RE digests of pRL1383a and BBa_C0012 on EtBr stained 2% agarose gel at 95V for 1 hour
- BBa_C0012 vector okay
- pRL1383a vector band ~9kb; no band observed at ~350bp. RE did not cut?
- Gel purified pRl1383a, BBa_C0012, GFP fusion, and Biobrick segments
- Used nanodrop spectrometer to determine DNA concentration (below)
- Ligated 7 μl Biobrick segment with 0.5 μl pRL1383a and 7 μl GFP fusion with 2 μl BBa_C0012 vector in 20 μl ligation reactions
- Transformed DH5α using 10 μl ligation reactions
- Plated GFP/C0012 vector on LB + amp100
- Plated BB segment/pRL1383a on LB + sp100
- Counted colonies from yesterday's pnir, slr2016, pilA assembly transformation (see experiment)
- Colony PCR'd colonies to verify presence of desired products
Transformation
Margaret
- Transformation of DB3.1 competent cells pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-) no plasmid, because transformation from 7/14/08 yielded only 1 colony from pSB3K3, nothing from pSB1A2 & pSB1AK3.
Plasmid prep
Krystle
- Resuspended yesterday's plasmid prep in 100 μl TE and stored in -20C
- Ran plasmid preps on gel to verify
- Grew up pnir, slr2016, and pilA transformants in LB in preparation for plasmid prep tomorrow
Restriction Digest
Krystle
- RE digested BBa_B0034, BBa_B0024, and BBa_E0040 with XbaI overnight
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 
]
