Team:Michigan/Oil Sands October
From 2010.igem.org
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**VF2- 33.9 nmol/.05 nmol/uL= 678 uL | **VF2- 33.9 nmol/.05 nmol/uL= 678 uL | ||
**VR- 40 nmol/ .05 nmol/uL= 800 uL | **VR- 40 nmol/ .05 nmol/uL= 800 uL | ||
- | *Used a 2 min | + | *Used a 2 min extension |
- | Also, put the Cm and Tet plates from Tuesday in the | + | Also, put the Cm and Tet plates from Tuesday in the 4ºC. |
== 10/23/2010 == | == 10/23/2010 == | ||
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'''Expected Digest bands''' | '''Expected Digest bands''' | ||
- | *FimB- 638 bp | + | *FimB - 638 bp |
- | *VP130- 2376, 1925 bp | + | *VP130 - 2376, 1925 bp |
- | *pSB1AT3-Flu- 6770, 15 bp | + | *pSB1AT3-Flu - 6770, 15 bp |
The digests appear to have worked, although the ligations seem to have been incomplete (Roche Rapid Dephos & Ligation Kit). | The digests appear to have worked, although the ligations seem to have been incomplete (Roche Rapid Dephos & Ligation Kit). | ||
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*16.7 uL UP H2O | *16.7 uL UP H2O | ||
- | We then did the following transformations using the [[Media:Direct Plating Transformation Protocol.pdf|Direct Plating Transformation Protocol]] and the | + | We then did the following transformations using the [[Media:Direct Plating Transformation Protocol.pdf|Direct Plating Transformation Protocol]] and the DH5α comp cells Nick made using his [[Media:Quick Transformation of E. coli Competant Cell Preperation.pdf| Competent Cell Preparation for Quick Transformation of E. coli]] protocol: |
Transformation (Plate Resistance): | Transformation (Plate Resistance): | ||
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Ligation: | Ligation: | ||
- | *pSB1C3- 4.572 fmol/uL | + | *pSB1C3 - 4.572 fmol/uL |
- | *VP130- 8.065 fmol/uL | + | *VP130 - 8.065 fmol/uL |
- | *FimB- 24.2 fmol/uL | + | *FimB - 24.2 fmol/uL |
VP130 Ligation Mix: | VP130 Ligation Mix: | ||
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Incubation: | Incubation: | ||
- | *10 mins @ 22. | + | *10 mins @ 22.5ºC |
- | *10 mins @ | + | *10 mins @ 65ºC |
Ligations were transformed according to [[Media:Direct Plating Transformation Protocol.pdf|Direct Plating Transformation Protocol]]. | Ligations were transformed according to [[Media:Direct Plating Transformation Protocol.pdf|Direct Plating Transformation Protocol]]. |
Latest revision as of 01:08, 27 October 2010