IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/15: Difference between revisions
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== | ==TesA/RBS Ligation Attempt 2== | ||
* | * Checked DNA concentrations | ||
:TesA= 11.99 ng/μL ; RBS=32.99 ng/μL | |||
*When divided by the number of base pairs: [TesA]=.018 and [RBS]=.016 | |||
* Ligation | * Ligation | ||
{| {{table}} border="1" align=center <!-- Ligation table --> | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent (μL) | |||
| bgcolor=#cfcfcf | Volume (μL) | |||
|- | |||
|| DNA(plasmid) || 1.0 μL RBS, 2.0 μL TesA | |||
|- | |||
|| 10X buffer || 1.0 | |||
|- | |||
|| dH<sub>2</sub>O || 5.0 | |||
|- | |||
|| Ligaze || 1.0 | |||
|- | |||
|| || 10.0 μL --> 25°C/ ~1 Hr. | |||
|} | |||
*After ligation, the mixture was heated to 65°C for 10 min. to denature the ligaze. | |||
*Transformation was then performed. | |||
:Three plates were grown overnight: RBS backbone (vector), TesA+RBS, and mCherryboth as a positive control and to replenish our stocks. | |||
* Transformation | * Transformation | ||
:Unfortunately, no growth was seen on any of the plates, including the positive control of mCherry. | |||
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Latest revision as of 00:06, 27 September 2017
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TesA/RBS Ligation Attempt 2
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