I have tried to isolate DNA from Padina but it is not getting amplified using markers that are working fine for other algal samples. Is there any specific method or precaution which should be followed?
Use Google/Pubmed to find out whether other researchers have had the same problem. Perhaps there is a known Taq Pol inhibitor in DNA preparations from that species? In that case, do serial dilutions of your sample and try them by PCR, to find out whether you can find a sweet spot at which the inhibitor is sufficiently diluted but the DNA is still amplifiable.
Please refer flowing links, they mention about methodology
https://www.google.lk/url?sa=t&rct=j&q=&esrc=s&source=web&cd=3&cad=rja&ved=0CC4QFjAC&url=http%3A%2F%2Fwww.researchgate.net%2Fpublication%2F227562500_An_effective_DNA_extraction_protocol_for_brown_algae%2Ffile%2F9fcfd50f616d6453bd.pdf&ei=Y7_pUraKCaSMiQf3noHICg&usg=AFQjCNG1tn6oRGRKlUrvpu4H6TA5mc195Q
https://www.google.lk/url?sa=t&rct=j&q=&esrc=s&source=web&cd=4&cad=rja&ved=0CDcQFjAD&url=http%3A%2F%2Fwww.vliz.be%2Fimisdocs%2Fpublications%2F98632.pdf&ei=Y7_pUraKCaSMiQf3noHICg&usg=AFQjCNFM4u5l9QcJKPdtxAtOzQkaiRKZyQ
http://www.oceannet.org/library/publications/documents/marine_samples.pdf
http://meacr.org/2ndACPrgnAbs.pdf
http://www.carsoncenter.uni-muenchen.de/download/publications/perspectives/2012_perspectives/1203_sickness_web_color.pdf
I found amplifications I did on shark tissue to be extremely sensitive to magnesium chloride concentrations. No idea if this will apply to Padina samples, but worth considering if the other suggestions don't help!
I have had a lot of problems in achieving PCR products from seaweed polysaccharide treated human faecal fermentation vessels. The only combination of extraction kit and Taq I was able to get working was the MO BIO PowerFaecal kit with KapaBiosystems Plant 3G Taq. I had to use quite a high MgCl2 conc with this combination as well. It should be noted that these problems were only apparent with certain seaweed species and not others.
- Badges
- Science method