Difference between revisions of "Kitchen-sink-ingredients"
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==Methylene blue rather than SYBRsafe== | ==Methylene blue rather than SYBRsafe== | ||
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| + | The first target was SYBR Safe.<ref>[https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/nucleic-acid-gel-electrophoresis/dna-stains/sybr-safe.html ThermoFischer's documentations for SYBR Safe]. Retrieved 27 August 2018.</ref> Even though SYBR Safe is far less toxic than Ethidium Bromide, SYBR Gold or other fluorescent dyes, it has not zero toxicity. By its mode of action (intercalation), it can be carcinogenic and the obligation to use a specific light (here a powerful blue light) is an obstacle for easy procedures and it can cause harm to the eyes of the user. <br> | ||
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| + | Therefore, we wanted to use Methylene Blue.<ref>[https://en.wikipedia.org/wiki/Methylene_blue Wikipedia page for Methylene Blue]. Retrieved 27 August 2018.</ref> As it is positively charged, it will bind to the outside structure of the DNA, without intercalating within it and without triggering much mutation. Plus, Methylene Blue is easy to find (it is on the WHO's list of essential medicines). Whereas for the Micronuclei Assay the methylene blue worked well, for the Comet Assay methylene blue did not give satisfactory results. <br> | ||
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| + | Photos of comet <br> | ||
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| + | ==References== | ||
| + | <references /> | ||
Latest revision as of 13:22, 4 September 2018
The most difficult objective is to find usual ingredients that can be easily found to replace the different chemicals needed for the experiment. We did not have enough time to study this precisely but still were able to compare the results of the experiment done in different conditions.
Sterile vs Bidistilled water
Methylene blue rather than SYBRsafe
The first target was SYBR Safe.<ref>ThermoFischer's documentations for SYBR Safe. Retrieved 27 August 2018.</ref> Even though SYBR Safe is far less toxic than Ethidium Bromide, SYBR Gold or other fluorescent dyes, it has not zero toxicity. By its mode of action (intercalation), it can be carcinogenic and the obligation to use a specific light (here a powerful blue light) is an obstacle for easy procedures and it can cause harm to the eyes of the user.
Therefore, we wanted to use Methylene Blue.<ref>Wikipedia page for Methylene Blue. Retrieved 27 August 2018.</ref> As it is positively charged, it will bind to the outside structure of the DNA, without intercalating within it and without triggering much mutation. Plus, Methylene Blue is easy to find (it is on the WHO's list of essential medicines). Whereas for the Micronuclei Assay the methylene blue worked well, for the Comet Assay methylene blue did not give satisfactory results.
Photos of comet
References
<references />