Have you been running a ladder along with each gel or some other positive control? That would definitively tell you whether the error is in the extraction/PCR or the running/staining of the gels. What size is the fragment that you are trying to amplify? From my experiences, one hour at 100V seems to be a fairly long time, but that might be the result of differences in fragment size that I have worked with and that you are trying to look at.
As for tests with foodstuffs, I remember doing a PCR experiment in high school to test for the presence of a sequence that is commonly part of GMO crops. Unfortunately, I do not remember any of the specifics of what the sequence was.
On Thursday, June 28, 2012 7:43:58 PM UTC-4, TRolandB wrote:
-- On Thursday, June 28, 2012 7:43:58 PM UTC-4, TRolandB wrote:
Hi there,The problem is that sometimes the procedure works and we get the correct bands showing up in electrophoresis, while other times we get no bands at all. We always follow the same general procedure, sometimes varying the quantities of reagents and PCR times to try and get it to work. Repeating the exact procedure from the times it did work doesn't lead to it working again.As the extraction process is the biggest variable we assume this is at fault. At the moment we extract it from cheek cells by scraping with a pipette tip. I've linked to the project page if anyone is interested, all info is there.CheersWill
On Thursday, 28 June 2012 13:59:39 UTC+1, phDIY wrote:Can you elaborate on your problems?
On Wednesday, June 27, 2012 6:12:10 PM UTC-5, TRolandB wrote:Hi everyone, our group (London hackspace) have been doing some genetic tests (sex typing) recently, with PCR and electrophoresis. We're getting very inconsistent results, and think our extraction process is at fault, so we want to try some similar techniques but with DNA from foodstuffs, so we can be sure of getting the extraction right.Does anyone have any suggestions of tests you can do that involve PCR and electrophoresis of this type?Will
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To view this discussion on the web visit https://groups.google.com/d/msg/diybio/-/qJHnZz7lluQJ.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
0 comments:
Post a Comment