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26/07/13
From 2013.igem.org
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| - | + | |Sample||Plasmid Volume(ul)||DNA concentration (ng/ul) | |
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| - | + | |1.1||40||131.8 | |
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| - | + | |1.2||40||77.1 | |
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| + | |1.3||46||56.8 | ||
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| + | |1.4||39||109 | ||
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| + | |1.5||41||129.1 | ||
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| + | |1.6||32||93.3 | ||
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*measured DNA concentration using nanodrop | *measured DNA concentration using nanodrop | ||
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==Restriction enzyme digest of the DNA samples== | ==Restriction enzyme digest of the DNA samples== | ||
Revision as of 13:16, 6 August 2013
| Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
|---|
| Sample | Plasmid Volume(ul) | DNA concentration (ng/ul) |
| 1.1 | 40 | 131.8 |
| 1.2 | 40 | 77.1 |
| 1.3 | 46 | 56.8 |
| 1.4 | 39 | 109 |
| 1.5 | 41 | 129.1 |
| 1.6 | 32 | 93.3 |
- measured DNA concentration using nanodrop
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
