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Clinical Microbiology & Molecular Biology Practices - Assignment Example

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In the paper “Clinical Microbiology & Molecular Biology Practices” the author discusses the agarose gel electrophoresis method, which depends on the agarose concentration, electrophoresis conditions, and the quality of the gel preparation to separate distinct fragments…
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Clinical Microbiology & Molecular Biology Practices
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Digested DNA samples are more likely to give better estimates of the  genome size than the undigested  DNA because smaller fragments resolve better giving distinct bands for individual fragments according to size followed by the calculation of the size of the individual bands from the standard curve which gives a more accurate size.
The agarose gel electrophoresis method depends on the agarose concentration, electrophoresis conditions, and the quality of the gel preparation to separate distinct fragments. Errors in the method could be due to distortion of bands produced as a result of a number of factors including migration of samples of the gel, loss of sample from the well due to leakage resulting in blurred bands, smearing of the bands due to loading too much DNA or it could even be due to the fact that agarose was not completely melted before the gel was poured. Errors could also be due to temperature and pH variation as well as cross-contamination of samples.
Generally,  DNA has five EcoRI restriction sites and HindIII has six. However, according to the results observed  DNA has four EcoRI restriction sites whereas HindIII has six. Literature shows contradictory statements on the number of HindIII digest of  DNA. However, it was found that wild type  DNA has six restriction sites for HindIII and standard lambda lab strain like cloud 1 ts857Sam7 has seven restriction sites as a result of single base substitution.
This does not agree with the digest of  EcoRI AND HindIII together. The linear  DNA genome generally shows six bands when digested with EcoRI and eight bands with HindIII. The discrepancy in the number of bands with EcoRI could be due to the errors in the method that might have resulted in the leakage of the sample. Read More
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