Project:Sex typing with amelogenin/results-20120301: Difference between revisions

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More photos of these results are available here: [[http://lardcave.net/biohacking/2012-03-02/]]
More photos of these results are available here: [[http://lardcave.net/biohacking/2012-03-02/]]
The descriptions below refer to gel lanes from the top downwards. Sorry, Rafael, but I didn't bother to rotate them this time :|


== Experiment 1 ==
== Experiment 1 ==
Visualised the previous night's PCR run on 1% agarose gel with 10ul/100ml ethidium bromide stock solution.
Visualised the previous night's PCR run on 1% agarose gel with 10ul/100ml ethidium bromide stock solution.


[[File:2012-02-29-results-1.JPG|200px|thumb|none|Results of experiment 1 (Nicholas, Helena, Mike, Tonderai)]]
[[File:2012-02-29-results-1.JPG|200px|thumb|none|Results of experiment 1 (Empty lane, ladder, Nicholas, Helena, Mike, Tonderai)]]


It is hard to see, but it almost looks like lane 4 (Tonderai? Or was it Mike?) is almost right -- there seem to be two distinct bands.
It is hard to see, but it almost looks like lane 4 (Tonderai? Or was it Mike?) is almost right -- there seem to be two distinct bands.
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I used the old master mix, as we have run out of the new stuff.
I used the old master mix, as we have run out of the new stuff.


[[File:2012-03-01-results-1.JPG|200px|thumb|none|Results of experiment 1 (Nicholas - meat tenderiser, Nicholas - meat tenderiser, Nicholas - heat to lyse)]]
[[File:2012-03-01-results-1.JPG|200px|thumb|none|Results of experiment 1 (Empty lane, ladder, Nicholas - meat tenderiser, Nicholas - meat tenderiser, Nicholas - heat to lyse)]]


Once again, two distinct bands are visible, but they don't seem to be in the right places on the gel.
Once again, two distinct bands are visible, but they don't seem to be in the right places on the gel.

Revision as of 21:20, 2 March 2012

More photos of these results are available here: [[1]]

The descriptions below refer to gel lanes from the top downwards. Sorry, Rafael, but I didn't bother to rotate them this time :|

Experiment 1

Visualised the previous night's PCR run on 1% agarose gel with 10ul/100ml ethidium bromide stock solution.

Results of experiment 1 (Empty lane, ladder, Nicholas, Helena, Mike, Tonderai)

It is hard to see, but it almost looks like lane 4 (Tonderai? Or was it Mike?) is almost right -- there seem to be two distinct bands.

Experiment 2

Nicholas ran another experiment during the day on March 1.

The protocol was as usual (see previous 4 or so notes), using the meat tenderiser method. He was careful to:

* Ensure the meat tenderiser / saliva solution was well mixed, with a 1-2 minute "sitting time" for the enzyme to take effect
* Ensure the alcohol was mixed with the tenderiser solution, in order to precipitate out the most DNA
* Ensure that the DNA, when extracted from the alcohol / saliva / tenderiser solution (yuck), was well dried, by heating for 5 min at 55 degrees.

I used the old master mix, as we have run out of the new stuff.

Results of experiment 1 (Empty lane, ladder, Nicholas - meat tenderiser, Nicholas - meat tenderiser, Nicholas - heat to lyse)

Once again, two distinct bands are visible, but they don't seem to be in the right places on the gel.

This is looking much more promising. It seems that it will now be a matter of fine tuning.

Also, the orange filter is amazing! Thanks to Stu.