Determination of purity, concentration and molecular weight of nucleic acids
- Clicks: 897 Release time: 2013/8/5
The mouse ELISA kit ethidium bromide is a fluorescent dye. In gel electrophoresis, EB molecules can be embedded between the base pairs of nucleic acid double strands and fluoresce under ultraviolet excitation, and the intensity is proportional to the amount of DNA. At the same time, the nucleic acid Zui has a large absorption wavelength of 260 nm. At this wavelength, the absorbance of 1 A corresponds to a certain concentration of nucleic acid, and the purity of the obtained nucleic acid can be determined by the ratio of A260/A280.
Experimental material DNA
Reagents, kits, agarose TBE, electrophoresis buffer, TE buffer, ethidium bromide (EB), sample buffer, instrument, electrophoresis, electrophoresis, electrophoresis, electronic, translation, nozzle, sample, microwave oven, ultraviolet spectrophotometer, gel image analyzer Experimental procedure
1. Turn on the spectrophotometer and warm up for 10 minutes.
2. Take 2 μl of the nucleic acid solution and add (or pure water) to make the volume 100 μl.
3. Add the diluted solution to the quartz tube and pour into another tube with TE buffer (or pure water) as standard.
4. Measure the absorbance at wavelengths 260 and 280 nm.
5. Compare readings at 260 nm and 280 nm.
Mouse ELISA kit considerations
1. A260/A280>1.8 indicates high purity of DNA/RNA. If the value is <<1.8, it means that the purity is low. At this time, the DNA content measured by OD260 is less reliable, and the nucleic acid solution contains more protein. Therefore, the nucleic acid obtained by the above is re-extracted.
2. Determine the absorbance at 260 nm. The value of OD=1.0 is as follows:
(1) ds DNA (double stranded DNA) = 50 mg/ml
(2) ss DNA (single stranded DNA or simgle-stranded RNA) = 40 mg/ml
(3) oligonucleotide = 33 mg/ml
3. Mouse ELISA kit EB is a carcinogen, do not touch with hands, not to pollute the environment.
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