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23/08/13
From 2013.igem.org
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Contents |
Nanodropping overnight incubations of herring sperm DNA
- Table representing the concentrations of herring sperm DNA before and after different treatments
| Sample | Concentration |
| Sheared/Unsonicated (s.u.s) | 17900ng/ul=17.9mg/ml |
| S.u.s 10fold dil. | 1790.1ng/ul |
| S.u.s 100fold dil | 152.6ng/ul |
| Sheared/Sonicated (s.s) | 17667ng/ul=17.6mg/ml |
| S.s 10fold dil. | 1766.7ng/ul |
| S.s 100fold dil. | 156.2ng/ul |
| Sheared/sonicated overnight | Concentration in the tube=Average of the two samples=16.9mg/ml |
| 500ul(DNA) in 500ul(1xTE) | 14485.9ng/ul |
| 2ml(DNA) in 10ml(1xTE) | 3883ng/ul |
- Concentrations decrease with dilutions
- The stock concentration in sheared/sonicated tube varies slightly, when derived from measured dilution concentrations that were subjected to different treatments (i.e. overnight incubation), though they orignated from the same tube
- Difference in concentration is not too significant though: proposed 17.6mg/ml vs 16.9mg/ml
Running overnight dilutions on gel
Lanes are as folllows: 1kb ladder, dilution 500ul DNA in 500 1xTE, dilution 2ml DNA in 10ml 1xTE, control unsheared/unsonicated 100 fold dilution.
Running 16s PCR of P. putida
Lanes are as follows: 100kb ladder, pcr samples 1, 2, 3, 4, 100kb ladder
Gel Extraction of PCR
- Cut out the bright band
- Use Thermo scientific geneJET extraction kit
