Difference between revisions of "Diy-transilluminator"
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Q5 polymerase was used for the amplification, and DNA from Sam (a non-taster) and Rachel (a taster) was used for the test. | Q5 polymerase was used for the amplification, and DNA from Sam (a non-taster) and Rachel (a taster) was used for the test. | ||
The DNA was a quick GnG prep that had been concentrated by pptn, after apparently no product was obtained the first PCR attempt, (but Rachel's sample had less DNA than Sam's... also less cells to start!). Stages of looking at the first gel are shown in images 6-8 of the gallery.<br> | The DNA was a quick GnG prep that had been concentrated by pptn, after apparently no product was obtained the first PCR attempt, (but Rachel's sample had less DNA than Sam's... also less cells to start!). Stages of looking at the first gel are shown in images 6-8 of the gallery.<br> | ||
| + | The new PCR with concentrated DNA was run on 11dec, and some was cut with HaeIII. Images from this expt are shown as 9-10 in gallery, but clear that this DIY transilluminator is not as sensitive as the UNIL 'SafeImager' from Promega - image 12... <br> | ||
The non-taster band that does not cut with HaeIII (predicted size 221bp) is visible from Sam's DNA, but no virtually no product came from Rachel's... (tasters are expected to have the band cut for 177 + 44bp perhaps there is a difference in the HaeIII cut lane of RA, vs the uncut, with something smaller perhaps just there??)<br> | The non-taster band that does not cut with HaeIII (predicted size 221bp) is visible from Sam's DNA, but no virtually no product came from Rachel's... (tasters are expected to have the band cut for 177 + 44bp perhaps there is a difference in the HaeIII cut lane of RA, vs the uncut, with something smaller perhaps just there??)<br> | ||
Revision as of 13:10, 12 December 2017
Transilluminators are especially useful to see DNA bands on a gel. While there is an old upcycled imager in the P1, this has detector problems, so sometimes much of the gel is not even visible... Therefore, we have been trying out new 'DIY-transilluminators' with a little help from some Hackteria friends and others (htgaa)!
DIY Transilluminator
Urs brought his GaudiLabs kit to Hackuarium on 2dec, and he helped Rachel do the soldering and put it all together.
He said it needs a diffuser of some sort to help avoid seeing the whole circuit board below.
The first five pictures in the gallery show various stages of this process.
11dec17 gel - second PTC PCR test
The chemical PTC can be tasted or not tasted, depending on a GC polymorphism in the TAS2R38 bitter taste receptor.
Primers for this test were obtained from Delphine Ducoulombier of l'Eprouvette.
Q5 polymerase was used for the amplification, and DNA from Sam (a non-taster) and Rachel (a taster) was used for the test.
The DNA was a quick GnG prep that had been concentrated by pptn, after apparently no product was obtained the first PCR attempt, (but Rachel's sample had less DNA than Sam's... also less cells to start!). Stages of looking at the first gel are shown in images 6-8 of the gallery.
The new PCR with concentrated DNA was run on 11dec, and some was cut with HaeIII. Images from this expt are shown as 9-10 in gallery, but clear that this DIY transilluminator is not as sensitive as the UNIL 'SafeImager' from Promega - image 12...
The non-taster band that does not cut with HaeIII (predicted size 221bp) is visible from Sam's DNA, but no virtually no product came from Rachel's... (tasters are expected to have the band cut for 177 + 44bp perhaps there is a difference in the HaeIII cut lane of RA, vs the uncut, with something smaller perhaps just there??)
Gallery
<gallery mode=packed widths=300 heights=300> File:IMG 2911.JPG File:IMG 2913.JPG File:IMG 2919.JPG File:IMG 2920.JPG File:IMG 2921.JPG File:IMG 3017.JPG File:IMG 3021.JPG File:IMG 3022.JPG File:IMG 3362.JPG File:IMG 3370.JPG File:1409taw4.jpg File:IMG 3373.JPG