Hi Ian,
Depending what type of DNA you ant to isolate. In case of bacterial genomic and plasmid DNA, just dilute a colony in buffer STET and boil it from 5 to 10 min at ~100ºC. Then, leave it getting cold on the bench until you see a clear phase at top ehere you can take sample for your PCR reaction. In case you don't have any of compounds for STET buffer, just try with DNAse-free water. I guess this will work for mammalian cells. Alternatively, try PCR from bacterial colony or cell cultures. I've made always massive clone checking using a small portion of bacterial colonies and this works pretty good..
Hi, I didn't try it but I found this: http://www.linkedin.com/company/a&a-biotechnology/micro-gravity-ax-dna-isolation-without-centrifugation-1682367/product
Cheers, Nadine
Hi,
you might find some magnetic beads to do the trick. There is a variety of beads for specific types of DNA, a quick google search showed. Good luck, cheers, Sebastian
If you have sufficient source material you can use the old-fashioned high-salt method and after precipitation just carefully fish out your DNA with a pipette. Do a google search and you will find detailed protocols. However, this will - as I mentioned - only work if you have really sufficient starting material.
Hi Ian,
Depending what type of DNA you ant to isolate. In case of bacterial genomic and plasmid DNA, just dilute a colony in buffer STET and boil it from 5 to 10 min at ~100ºC. Then, leave it getting cold on the bench until you see a clear phase at top ehere you can take sample for your PCR reaction. In case you don't have any of compounds for STET buffer, just try with DNAse-free water. I guess this will work for mammalian cells. Alternatively, try PCR from bacterial colony or cell cultures. I've made always massive clone checking using a small portion of bacterial colonies and this works pretty good..
You could make yourself a Dremelfuge for under $20. It's easy and fun. Search it on Google. (and wear a face shield ;-)
yes colony pcr could help wherein you may lyse the cell in a buffer and use it directly for your reaction. the lysis conditions for Gram -ve and +ve organisms are different (incase you are working with bacterial cells).
a fresh colony is always preferred over an old one to ensure maximum DNA recovery and it also minimizes the changes of ending up with degraded DNA
well well............i suggest a fantastic method for it......extraction of DNA from different biological samples using magnetic nanoparticles.....process will take less then an hour and the yield will be comparable or higher than any other method.......
A general method is to get a TINY bit of material on the end of a toothpick, put it into a PCR tube and microwave it for a while (I do 90s full power for about 10-15 samples of fungal conidia on wet sterile toothpicks). Then just add your PCR mix onto the toothpick, mix it up and remove the pick. Close lid and put into the PCR machine. But you may have to be careful which polymerase you use; I use Phusion from NEB as standard Taq doesn't seem to like this. If it works for your material it does save a great deal of work.
There are also column systems that use vacuum to remove solutions
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