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23/08/13
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| - | ==Nanodropping overnight incubations of herring sperm== | + | <div style="font-family:arial;padding:5px;border-radius:5px;border:5px solid #FF2800;"> |
| - | *Table representing the concentrations of herring sperm | + | {| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border="0" bordercolor="#000000" width="100%" align="center" |
| + | !align="center"|[[Team:Leicester|Home]] | ||
| + | !align="center"|[[Team:Leicester/Team|Team]] | ||
| + | !align="center"|[https://igem.org/Team.cgi?year=2013&team_name=Leicester Official Team Profile] | ||
| + | !align="center"|[[Team:Leicester/Project|Project]] | ||
| + | !align="center"|[[Team:Leicester/Parts|Parts Submitted to the Registry]] | ||
| + | !align="center"|[[Team:Leicester/Modeling|Modeling]] | ||
| + | !align="center"|[[Team:Leicester/Notebook|Notebook]] | ||
| + | !align="center"|[[Team:Leicester/Safety|Safety]] | ||
| + | !align="center"|[[Team:Leicester/Attributions|Attributions]] | ||
| + | |} | ||
| + | </div> | ||
| + | |||
| + | ==Nanodropping overnight incubations of herring sperm DNA== | ||
| + | *Table representing the concentrations of herring sperm DNA before and after different treatments | ||
{|border=1 | {|border=1 | ||
|Sample||Concentration | |Sample||Concentration | ||
|- | |- | ||
| - | |Sheared/Unsonicated (s.u.s)||17900ng/ul=17.9mg/ml | + | |'''Sheared/Unsonicated''' (s.u.s)||17900ng/ul='''17.9mg/ml''' |
|- | |- | ||
|S.u.s 10fold dil.||1790.1ng/ul | |S.u.s 10fold dil.||1790.1ng/ul | ||
| Line 10: | Line 24: | ||
|S.u.s 100fold dil||152.6ng/ul | |S.u.s 100fold dil||152.6ng/ul | ||
|- | |- | ||
| - | |Sheared/Sonicated (s.s)||17667ng/ul=17.6mg/ml | + | |'''Sheared/Sonicated (s.s)'''||17667ng/ul='''17.6mg/ml''' |
|- | |- | ||
|S.s 10fold dil.||1766.7ng/ul | |S.s 10fold dil.||1766.7ng/ul | ||
| Line 16: | Line 30: | ||
|S.s 100fold dil.||156.2ng/ul | |S.s 100fold dil.||156.2ng/ul | ||
|- | |- | ||
| - | |Sheared/sonicated overnight||Concentration in the tube=Average of the two samples=16.9mg/ml | + | |'''Sheared/sonicated overnight'''||Concentration in the tube=Average of the two samples='''16.9mg/ml''' |
|- | |- | ||
| - | |500ul | + | |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== | ||
| + | [[File: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''== | ||
| + | |||
| + | [[File: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 | ||
Latest revision as of 11:44, 27 August 2013
| Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
|---|
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
