DNA Isolation Report Summary Table Final Volume of DNA Preparation (mL) 2 A 260 0.521 A 280 0.252 A 260 ~A 280 2.0675 Purity Purity >2 possible RNA contamination Dilution factor used for UV spectrum 1:20 Diluted [DNA] (μg/mL) 9.18 Original [DNA] (mg/mL) 0.184 Yield (mg)* 0.367 Yield (mg/g wet wt cells) 0.1835 Expected Yield (mg) based on the theoretical calculations 0.734 The DNA Spectrum Method A culture of E.coli (~0.1g wet weight per mL), suspended in 0.1 M NaCl, 10 mM EDTA, pH 8. Incubated with a 2% w/v SDS for 10 minutes at 65 degrees. Following the incubation, Sodium Perchlorate was added to 1mM and the solution was extracted with an equal volume of chloroform: amyl alcohol (24:1) solution. After shaking for 20 minutes in a mechanical shaker at room temperature, remove the upper aqueous layer. Layer 2 volumes of 100% cold ethanol carefully on top of the solution. Collect the nucleic acid by spooling. The precippitate (nucleic acids) is then washed briefly in 70% (v/v) ethanol and then is dissolved in 10mM Tris and
DNA Isolation Report
Jan 04, 2016
Report and procedure on isolating DNA from a culture of e.Coli bacteria
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DNA Isolation Report
Summary Table
Final Volume of DNA Preparation (mL) 2A260 0.521A280 0.252A260~A280 2.0675Purity Purity >2 possible RNA contaminationDilution factor used for UV spectrum 1:20Diluted [DNA] (μg/mL) 9.18Original [DNA] (mg/mL) 0.184Yield (mg)* 0.367
Yield (mg/g wet wt cells) 0.1835Expected Yield (mg) based on the theoretical calculations
0.734
The DNA Spectrum
MethodA culture of E.coli (~0.1g wet weight per mL), suspended in 0.1 M NaCl, 10 mM EDTA, pH 8. Incubated with a 2% w/v SDS for 10 minutes at 65 degrees. Following the incubation, Sodium Perchlorate was added to 1mM and the solution was extracted with an equal volume of chloroform: amyl alcohol (24:1) solution. After shaking for 20 minutes in a mechanical shaker at room temperature, remove the upper aqueous layer. Layer 2 volumes of 100% cold ethanol carefully on top of the solution. Collect the nucleic acid by spooling. The precippitate (nucleic acids) is then washed briefly in 70% (v/v) ethanol and then is dissolved in 10mM Tris and 1 mM EDTA (0.4 ml/mL of E.coli suspension). The quality and yield was assessed by the UV spectrophotometry and by gel electrophoresis and without RNAse treatment.
ExplanationThese changes would not alter the method because it uses the same concentrations to isolate and extract the DNA. However the values for SDS and the amount of DNA in the starting solution would change due to different however by altering the volumes used gives back a constant concentration to isolate the DNA.
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