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Discussion

DNA bands and its quality


After electrophoresis at 103 to 104 volts for 10 minutes, the DNA bands were
observed under ultra-violent light and the images were saved in a gel analyser .
The quality of DNA bands for A and B samples that’s the fours last bands do not
accurately show the quantity of the base pairing present. As it can be seen, the
picture contains several bands with different length since sample A and B may
contains different level of contaminates and DNA. The longer are the bands the
higher is the DNA concentration. When electrophoresis are being conducted,
smaller DNA fragments travels at a faster rate and larger DNA fragments travel at
lower rate. The bands can be also distinguishable from each other. However the
majority of the bands appear quit pale and less fluorescent, this can be due to the
lower percentage of agarose gel which was used such as 0.4% and the absence of a
ladder during electrophoresis. Most agarose gels are made with between 0.7%
providing good separation or resolution of large 5-10kb DNA fragments and 2%
for good resolution for small 0.2-1kb fragments and the agarose dissolved in
electrophoresis buffer is 1% gels which are commonly used for many applications.
Another error might have been the bubbles created when pouring agarose gel
into the comb. In order to improve visualization quality and the concentration of
DNA sample, the use of SYBR Green I or SYBR is better instead of ethidium
bromide which is 25 times more sensitive and very less mutagenic.
Spectrophotometer and DNA extraction

Spectrophotometer was used to measure the absorbance of A and B molluscan


sample. The values of absorbance (A) and (B) allowed estimating the purity,
concentration and yield of the genomic DNA samples.
The values of sample A and B exhibited the A260/ A280 ratio were 1.354 and
1.407 respectively which is below the range of 1.8 – 2.0 for pure DNA. Sample A
and B contains molluscan DNA which is difficult to obtain high purity level of
DNA .Firstly because of the large amount of mucopolysaccharides and
polyphenolic proteins in their tissues and secondly genetic studies have shown in
the scarcity of tissue in mollusc therefore these affects the isolation of DNA
samples. The presence of these contaminants cause damage of the DNA and/or
inhibit enzymatic reactions. High level of sugar, protein contamination cause
interference in downstream molecular biology procedures. . The level of DNA
concentrations which were calculated for A was 1.0725mg/ml whereas B is 0.8745
mg/ml. The analyses shows that sample A contains higher level of DNA .This can
be due to the difference in their tissue contents. Quantity and quality of the isolated
DNA can be improved by using manual extraction than automatic extraction
methods
https://www.researchgate.net/publication/51884609_An_Efficient_Method_for_Genomic_DNA_Extract
ion_from_Different_Molluscs_Species

https://www.researchgate.net/publication/305401989_DNA_analysis_of_molluscs_from_a_museum_w
et_collection_A_comparison_of_different_extraction_methods

https://www.researchgate.net/post/Why_I_cant_see_any_DNA_band_on_Agarose_gel

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