DNA protocols: from field to sequencer
24-04-2009 21:27
Preparation of material for DNA research is becoming
more and more important, as the facilities are
continuously improving. At Naturalis we will make a
‘quantum leap’ later this year when a new molecular
lab will be opened.
Recently there were some suggestions made by colleagues that the procedure I used so far, might not be full-proof for DNA. Especially the use of 'alcohol-proof' paper for labeling was not recommended by an expert, since this paper contains a very low concentration of formaldehyde (which in high concentrations destroys DNA).
Also there are some bad experiences mentioned with the niku-nuki method when applied to larger snails (the method was developed for smaller ones). I need to do some further testing on that and see how we may optimize the method for animals of orthalicid size (often 2+ cm).
Thirdly there was the suggestion to take tissue samples from living material, either from the foot or the rim of the mantle. This could be done with an extreme sharp knife and it was suggested that the snails could survive when the sample was small enough.
Finally, a colleague mentioned Nembutal, a substance I used in the past to stretch the snails before killing them in alcohol.
This are clearly things that need more elaboration. An inventory of pros and cons of different protocols for the preparation of animals could be helpful. What works best for you? What is a practical method when you are out in the field? What if you have animals in the lab? Do you have good results with a particular protocol geared towards larger snails?
I would be interested to hear your comments on this issue.
Recently there were some suggestions made by colleagues that the procedure I used so far, might not be full-proof for DNA. Especially the use of 'alcohol-proof' paper for labeling was not recommended by an expert, since this paper contains a very low concentration of formaldehyde (which in high concentrations destroys DNA).
Also there are some bad experiences mentioned with the niku-nuki method when applied to larger snails (the method was developed for smaller ones). I need to do some further testing on that and see how we may optimize the method for animals of orthalicid size (often 2+ cm).
Thirdly there was the suggestion to take tissue samples from living material, either from the foot or the rim of the mantle. This could be done with an extreme sharp knife and it was suggested that the snails could survive when the sample was small enough.
Finally, a colleague mentioned Nembutal, a substance I used in the past to stretch the snails before killing them in alcohol.
This are clearly things that need more elaboration. An inventory of pros and cons of different protocols for the preparation of animals could be helpful. What works best for you? What is a practical method when you are out in the field? What if you have animals in the lab? Do you have good results with a particular protocol geared towards larger snails?
I would be interested to hear your comments on this issue.