User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/06: Difference between revisions
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==01/06/13== | ==01/06/13== | ||
<!-- Precede finished items with a checkmark ✓ --> | <!-- Precede finished items with a checkmark ✓ --> | ||
* Gibson | * Gibson Assembly: pick colonies & check via colony PCR | ||
* Media: Make LB agar +amp | * Media: Make LB agar +amp | ||
* Gibson Assembly: redo transformation with more DNA | |||
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| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| bgcolor=#cfcfcf | 5x mix | | bgcolor=#cfcfcf | 5x mix | ||
| rowspan="7" | [[Image:KAH010613_gel1.jpg|270px|PCR gel]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |- | ||
| Colony || --- || --- | | Colony || --- || --- | ||
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* 4°C, ∞ | * 4°C, ∞ | ||
Conclusion: looks like primer dimer. Re-do the transformation with 3x more DNA (from yesterday, stored at -20°C) | |||
---- | ---- | ||
''' | '''Transformation''' | ||
# | # hPCD + BL01 + V0120, 6.0 μL 1/4x Gibson | ||
# | # V0120, 6.0 μL 1/4x Gibson | ||
# hPCD + BL01 + pSB1A3, 6.0 μL 1/4x Gibson | |||
# pSB1A3, 6.0 μL 1/4x Gibson | |||
* Add DNA to 50 μL DH5α-Turbo; ice/ 10 min. | |||
* Plate on 100 μg/mL amp. | |||
Revision as of 17:38, 6 January 2013
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01/06/13
Gibson Assembly results
PCR
Conclusion: looks like primer dimer. Re-do the transformation with 3x more DNA (from yesterday, stored at -20°C)
Transformation
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