IGEM:Peking University/2008/Experiment/PKU 0620 G2 1: Difference between revisions
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::'''Enzyme cutting- | ::'''Enzyme cutting-pPT1-XbaI-20 μL''' | ||
{|border="1" cellspacing="0" | {|border="1" cellspacing="0" | ||
|+ | |+ | ||
| Line 61: | Line 59: | ||
| ||Volumn(μL) | | ||Volumn(μL) | ||
|- | |- | ||
| | | plasmid||5 | ||
|- | |- | ||
| | | XbaI||0.2 | ||
|- | |- | ||
| | | BSA ||2 | ||
|- | |- | ||
| M Buffer||2 | | M Buffer||2 | ||
|- | |- | ||
|ddH<sub>2</sub>O|| | |ddH<sub>2</sub>O||10.8 | ||
|} | |} | ||
{|align="right" | {|align="right" | ||
|back to [[PKU_Experiment|Previous Page]] | |back to [[PKU_Experiment|Previous Page]] | ||
|} | |} | ||
Revision as of 05:22, 4 July 2008
Primer design
- Primer 1
- Primer name: F-HindIII-Apal-Gal4
- Sequence(5'to3'):CCCAAGCTTGGGCCCAAGATGAAGCTACTGTCTTCTATCGAACAA
- Tm:64.9°C
- Primer 2
- Primer name: R-EcoRI-Spel-Gal4
- Sequence(5'to3'):GGGGAATTCACTAGTATTTTACTCTTTTTTTGGGTTTGGTGGGGT
- Tm:61.3°C
PCR
| Volume (μL) | |
| Template （pPT1） | 0.5 |
| dNTP | 1 |
| Enzyme | 0.5 |
| Buffer | 10 |
| F-HindIII-Apal-Gal4 | 0.5 |
| R-EcoRI-Spel-Gal4 | 0.5 |
| ddH2O | 37 |
- conditions
| Temperature | Time Length(0:00:00) | |
| 98°C | 00:30 | |
| 98°C | 00:10 | 30 cycles |
| 57.1,60.4,62.5,64.7,66.9,68.9°C | 00:30 | 30 cycles |
| 72°C | 01:30 | 30 cycles |
| 72°C | 07:00 | |
| 4°C | Hold |
- Enzyme cutting-pPT1-XbaI-20 μL
| Volumn(μL) | |
| plasmid | 5 |
| XbaI | 0.2 |
| BSA | 2 |
| M Buffer | 2 |
| ddH2O | 10.8 |
| back to Previous Page |
