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26/07/13
From 2013.igem.org
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!align="center"|[[Team:Leicester|Home]] | !align="center"|[[Team:Leicester|Home]] | ||
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==Purification of the limonene biobrick - BBa_24== | ==Purification of the limonene biobrick - BBa_24== | ||
Revision as of 13:56, 1 August 2013
| Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
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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 then 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 5ul of ethidium bromide.
- Pour and let it set for 30 mins.
- Adding dye and marker
- Used orange g loading dye.
- 2ul to each sample
- Used 1kb ladder, 5ul per 1 marker well.
- Run the gel
- Gel picture
