Toronto/Lab Protocols/Quantitation
From 2007.igem.org
< Toronto | Lab Protocols
Quantitation
- Take the extracted DNA and run a gel to find out the concentration (per μL) of your DNA.
- 1 μL plasmid + 4 μL ddH2O + 1 μL Loading Dye
- 1 μL insert + 4 μL ddH2O + 1 μL Loading Dye
Gel Lane | Fragment |
---|---|
1 | HindIII Ladder (5 μL) |
2 | HindIII Ladder (2 μL) |
3 | HindIII Ladder (1 μL) |
4 | Blank |
5 | Plasmid (All of it) |
6 | Blank |
7 | Insert (All of it) |
8 | Blank |
9 | 1 Kbp Ladder (3 μL) |
- Ensure that the plasmid and inserts are at the correct lengths using the 1 Kbp ladder.
- Note that 2 μL of HindIII will twice as bright as 1 μL of HindIII, and 5 μL of HindIII will be five times as bright. So to determine concentration, estimate the intensity of the bands compared with the 3 HindIII ladders and assign it a relative intensity factor. Read off the concentration value on the Ladder card, and multiply by the relative intensity factor to determine your sample concentration in ng/μL.
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