Dear group members,
I ve 2bacterial cells dt can degrade crude oil synergistically,after plasmid curing they failed 2 degrade.wot protocol can I use 2identify rls coded in d plasmid dt exist btw them
hi i think your cells are producing biosurfactant can you tell me the microbes name? have u tested the extra cellular extract of bacteria for oil degradation
Hi Ajao,
By all mean, I think you would like to know about "plasmid extraction, purification and sequencing protocols". But I do hope you basically have known your bacteria at lease at family and genus level. In addition, I guess you have some idea in mind about oil degrading functions that encoded on the plasmids of your bacteria (encoding enzymes or biosynthetic molecules that involve and effect oil degradation). If it is so, there are many commercial plasmid extraction and purification prep. prior sequencing submission (such as QIAGEN) . However, I suggest couple protocols (out of many) for your own preparation. I do concern about the targeted plasmids before sequencing. I recommend targeting the plasmids from obtained info; bacterial genus specie, functioning character of oil degradation, then size of plasmid, etc. compared to plasmid library or plasmid data base.
References;
Plasmid extraction & purification
[1] https://tspace.library.utoronto.ca/bitstream/1807/1076/1/jb03018.pdf
[2] http://www.ncbi.nlm.nih.gov/pmc/articles/PMC330384/pdf/nar00188-0356.pdf
[3] http://www.protocol-online.org/prot/Protocols/Isolation-of-Plasmid-DNA-3922.html
Plasmid library (database)
[1] http://plasmid.med.harvard.edu/PLASMID/PrepareBlast.do
[2] http://gind-db.ucsf.edu:8000/cgi-bin/Plasmid/plasmid_search2.cgi
Hope these info may be a little helper.
Cheers
God bless you but I will still have to disturb you if need be!!!!!!!!!!!!
Can you please suggest any lab. where 16SrDNA can be run for proper identification of my isolates before any further work.Please you can kindly send it to my inbox [email protected]
Hi Ajao,
Sorry for late reply as I have been busy. I sent you the information to inbox of your account of ResearchGate. Check it out and good luck. Cheers,
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