User:Jose Fabricio Lopez/Notebook/Logbook/2011/09/11: Difference between revisions
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(Autocreate 2011/09/11 Entry for User:Jose_Fabricio_Lopez/Notebook/Logbook) |
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== | =Goals= | ||
* Repeat Backbones for these [[User:Jose Fabricio Lopez/Notebook/Logbook/2011/09/08|PCRs]] | |||
==PCR== | |||
{|border=1| | |||
|+ 3 PCRs for TerB0015-Backbone | |||
<br> | |||
Oligos for 1st Terminator | |||
!Reagents | |||
! μL | |||
!MegaMix | |||
|-- | |||
|H<sub>2</sub>O | |||
|24.5 | |||
|74 | |||
|-- | |||
|5X Phusion Buffer HF | |||
|10 | |||
|30.5 | |||
|-- | |||
|10mM dNTPs | |||
|1 each | |||
|3.1 each | |||
|-- | |||
|Primer F | |||
|5 | |||
|15.2 | |||
|-- | |||
|Primer R | |||
|5 | |||
|15.2 | |||
|-- | |||
|DNA | |||
|1 | |||
|3 | |||
|-- | |||
|Phusion | |||
|.5 | |||
|NA | |||
|} | |||
{|border=1| | |||
|+ 3 PCR for TerB0015-Backbone | |||
<br> | |||
Oligos for 2nd Terminator | |||
!Reagents | |||
! μL | |||
!MegaMix | |||
|-- | |||
|H<sub>2</sub>O | |||
|24.5 | |||
|74 | |||
|-- | |||
|5X Phusion Buffer HF | |||
|10 | |||
|30.5 | |||
|-- | |||
|10mM dNTPs | |||
|1 each | |||
|3.1 each | |||
|-- | |||
|Primer F | |||
|5 | |||
|15.2 | |||
|-- | |||
|Primer R | |||
|5 | |||
|15.2 | |||
|-- | |||
|DNA | |||
|1 | |||
|3 | |||
|-- | |||
|Phusion | |||
|.5 | |||
|NA | |||
|} | |||
{|border="1" | |||
!Cycles | |||
!Gel instruct. | |||
!Gel | |||
|- | |||
| | |||
*98°C 30s | |||
*98°C 10s 30Cy | |||
*60°C 30S 30Cy | |||
*72°C 1:50 30Cy | |||
*72°C 10min | |||
*4°C | |||
| | |||
*Lane2 Ladder | |||
*Lane3-5 Ter1-Backbone 1 | |||
*Lane6-8 Ter2-Backbone 2 | |||
|[[Image:PCRBothTerminators.jpg|200px]] | |||
|} | |||
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__NOTOC__ | __NOTOC__ | ||
Revision as of 15:06, 12 September 2011
 iGEM 2011
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Goals
PCR
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