Project:Sex typing with amelogenin/results-20120301

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. This is twice the amount of EtBr as usual, but it doesn't make a difference (see below).

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.

Visualised on 1% agarose gel with 6ul/100ml ethidium bromide solution. NB we have previously been using 5ul/100ml. Based on these results I am not convinced that more EtBr is at all helpful -- I noticed no difference in brightness between these two runs.

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.