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26/07/13
From 2013.igem.org
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| - | + | purification of the limonene biobrick - BBa_24 | |
| - | * | + | *Plasmid Volume |
| - | + | -1.1 - 40ul | |
| - | * | + | -1.2 - 40ul |
| - | * | + | -1.3 - 46ul |
| + | -1.4 - 39ul | ||
| + | -1.5 - 41ul | ||
| + | -1.2 - 32ul | ||
| + | *measured DNA concentration using nanodrop | ||
| + | *DNA concentration (ng) | ||
| + | -1.1 - 131.8 | ||
| + | -1.2 - 77.1 | ||
| + | -1.3 - 56.8 | ||
| + | -1.4 - 109 | ||
| + | -1.5 - 129.1 | ||
| + | -1.2 - 93.3 | ||
| - | + | *Restriction enzyme digest of the DNA samples | |
| - | + | -enzymes: XbaI, PstI | |
| + | -master mix: Buffer - 14ul | ||
| + | H2O - 88.2ul | ||
| + | XbaI - 1.4ul | ||
| + | PstI - 1.4ul | ||
| + | Total volume - 105ul | ||
| + | Each sample contained 5ul of DNA and 15ul of the master mix. | ||
| + | The total amount of DNA in each sample was 200ng | ||
| + | To calculate it we divided 200ng by the concentration of each sample | ||
| + | The rest of the volume was made up with water to give a total of 5ul. | ||
| + | The samples were them incubated in a water bath at 37C by an hour | ||
| + | |||
| + | *Agorose Gel preparation | ||
| + | -20ml of 5xTBE | ||
| + | -80ml of water | ||
| + | -0.8g of agarose | ||
| + | |||
| + | Boil for 3 minutes, 1 minute at a time. | ||
| + | Let it cool and add 5um of ethidium bromide. | ||
| + | Pour and let set for 30 mins. | ||
| + | |||
| + | *Adding dye and and marker | ||
| + | Used orange g loading dye. | ||
| + | -2um to each sample | ||
| + | |||
| + | Used 1kb ladder, 5um per 1 marker well. | ||
| + | |||
| + | *Run the gel | ||
Revision as of 15:36, 26 July 2013
purification of the limonene biobrick - BBa_24
- Plasmid Volume
-1.1 - 40ul -1.2 - 40ul -1.3 - 46ul -1.4 - 39ul -1.5 - 41ul -1.2 - 32ul
- measured DNA concentration using nanodrop
- DNA concentration (ng)
-1.1 - 131.8 -1.2 - 77.1 -1.3 - 56.8 -1.4 - 109 -1.5 - 129.1 -1.2 - 93.3
- Restriction enzyme digest of the DNA samples
-enzymes: XbaI, PstI -master mix: Buffer - 14ul
H2O - 88.2ul
XbaI - 1.4ul
PstI - 1.4ul
Total volume - 105ul
Each sample contained 5ul of DNA and 15ul of the master mix. The total amount of DNA in each sample was 200ng To calculate it we divided 200ng by the concentration of each sample The rest of the volume was made up with water to give a total of 5ul. The samples were them incubated in a water bath at 37C by an hour
- Agorose Gel preparation
-20ml of 5xTBE -80ml of water -0.8g of agarose
Boil for 3 minutes, 1 minute at a time. Let it cool and add 5um of ethidium bromide. Pour and let set for 30 mins.
- Adding dye and and marker
Used orange g loading dye. -2um to each sample
Used 1kb ladder, 5um per 1 marker well.
- Run the gel
