23/08/13

From 2013.igem.org

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Nanodropping overnight incubations of herring sperm DNA

  • Table representing the concentrations of herring sperm DNA before and after different treatments
SampleConcentration
Sheared/Unsonicated (s.u.s)17900ng/ul=17.9mg/ml
S.u.s 10fold dil.1790.1ng/ul
S.u.s 100fold dil152.6ng/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 overnightConcentration 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

Igem ovenight incub herrinngDNA 230813.jpeg


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

Igem PCR 230813.jpeg


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