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| <h5 style="color:indigo" id="spectrophotometer">Nanodrop analysis </h5> | | <h5 style="color:indigo" id="spectrophotometer">Nanodrop analysis </h5> |
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− | <img src="https://static.igem.org/mediawiki/2019/d/de/T--EPFL--PCR_MN_EC.jpg" >
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| <div class="post-content"><p style="font-size:17px;" align="justify"> | | <div class="post-content"><p style="font-size:17px;" align="justify"> |
− | We used the Nanodrop to analyze the DNA extracted with a microneedle patch (red). The positive control (green) | + | We used the Nanodrop to analyze the DNA extracted with a microneedle patch (red). The positive control (green) is EC synthetic DNA sequence and the negative control (blue) is the elution buffer applied on an unused microneedle patch. </br> |
− | is EC synthetic DNA sequence and the negative control (blue) is the elution buffer applied on an unused microneedle patch. </br>
| + | As expected, the negative control absorption at 260nm is close to zero, no DNA is present in the microneedle patch. In contrast, the positive control has the typical DNA Nanodrop readout. </br> |
− | As expected, the negative control absorption at 260nm is close to zero, no DNA is present in the microneedle patch.
| + | The graph of DNA extracted by microneedle patch shows a peak of absorption at 260nm, we can also see that that values absorption values at 230nm and 280nm are lower that the value at 260nm. Overall, the graph is similar to the positive control but with lower values. This indicates that DNA was extracted by the microneedle patch. |
− | In contrast, the positive control has the typical DNA Nanodrop readout. </br>
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− | The DNA extracted by microneedle patch shows significant absorption at A260. Its line higher than the negative
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− | control but lower than the positive control. This means that some DNA was extracted but that it is not pure,
| + | <img src="https://static.igem.org/mediawiki/2019/2/22/T--EPFL--courbes.png" > |
− | which is normal since, as mentioned previously, the patch extracts DNA and other molecules on the leaf.
| + | </br> </p> |
− | </p>
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| <h5 style="color:indigo" id="Gel">PCR amplification</h5> | | <h5 style="color:indigo" id="Gel">PCR amplification</h5> |
− | <img src="https://static.igem.org/mediawiki/2019/c/c9/T--EPFL--3courbes.png" >
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| <p style="font-size:17px; align="justify"> | | <p style="font-size:17px; align="justify"> |
| We amplified different EC DNA by PCR and ran a gel electrophoresis to analyze the results. | | We amplified different EC DNA by PCR and ran a gel electrophoresis to analyze the results. |
− | We loaded the gel from left to right as following, the ladder, synthetic EC DNA sequence,
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− | DNA extracted with a microneedle patch, DNA extracted with a standard kit, control with primers but no DNA.
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| Bands are observed in all the lanes containing DNA, no band is present in the control. | | Bands are observed in all the lanes containing DNA, no band is present in the control. |
| This shows that the microneedle patch successfully extracted DNA from the plant, which was then amplified by PCR. | | This shows that the microneedle patch successfully extracted DNA from the plant, which was then amplified by PCR. |
| </p> | | </p> |
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− | | + | <img src="https://static.igem.org/mediawiki/2019/d/de/T--EPFL--PCR_MN_EC.jpg" > |
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