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Posted

hello.

I need help in finding a primer for my DNA sample. i need a primer for iron reducing bacteria gene. I've tried to find in over the internet but couldn't find it. Anyone care to help?

Posted
hello.

i need a primer for iron reducing bacteria gene.

 

Can you not design it yourself? I mean design one from the sequence of whatever bit you want and then order it from a company that makes oligonucleotides? Isn't that what most labs do? I take it you're a student - ask where other people get their primers from?

Posted

thanx...

i am doing PCR and i try to amplify this sequence to prove that the bacteria contain the iron reducing gene. The problem is, i don't know the primer used to amplify the target sequence. I want to use a specific primer, not the 16s rRNA

by the way, thanx a lot

Posted

I did a bit more reading on the subject (I'm interested in bacterial metabolism anyway) and I've reached the conclusion that there is no such thing as THE gene for iron reducing. The iron-reducing bacteria are a very diverse group and only two organisms, Geobacter and Shewanella, have been studied in detail.

Here's two examples of relevant studies of Geobacter:

Combining in-situ reverse transcriptase polymerase chain reaction, microscopy and x-ray photoelectron microscopy to investigate mineral surface-associated microbial activities

OmcF, a Putative c-Type Monoheme Outer Membrane Cytochrome Required for the Expression of Other Outer Membrane Cytochromes in Geobacter sulfurreducens

Studies to find evidence for, and characterisation of, iron-reducing bacteria seems to have been undertaken by 16s rRNS analysis, though some have been using more specific primers than the general bacterial ones. For example, I came across this article, Diversity of Geobacteraceae Species Inhabiting Metal-Polluted Freshwater Lake Sediments Ascertained by 16S rDNA Analyses, which uses a Geobacteraceae-specific primer. Of course, you would be missing out on the other iron-reducing groups this way.

 

I'm afraid this is not really the answer you're looking for. But I hope that you will find a good way to proceed with your work.

 

Airmid.

Posted

In order to PCR amplify your putative iron reducing gene, you need to figure out the sequence flanking the gene in the E. coli genome. Using that sequence information, you can design sequences completmentary to the region upstream of the gene on the antisense strand and complementary to the downstream region of the sense strand. In general, PCR primers must be designed by the researcher and are not found by searching on the internet.

Posted

thanx for the help. I think, it should be good enough for me to think of something for my work. Really appreciate it.

I thought, there is somebody else who already design the primer, so i can find it through the internet, but i couldn't find one. by the way, thanx again.

Posted
hello.

I need help in finding a primer for my DNA sample. i need a primer for iron reducing bacteria gene. I've tried to find in over the internet but couldn't find it. Anyone care to help?

 

An iron reducing gene? huh? What does that mean? Am I being stupid?

Posted

Nashyboyo, my DNA sample contains gene that is able to reduce the iron compound. And, what i am doing rite now is to detect that gene by using the PCR. but the prblem is, i don't have the specific primer to amplify it. So, i thought, maybe, there is someone here could suggest any site or paper that i can use for my study.

Thank you.

  • 2 weeks later...
Posted

thanx for all the help...

i have another question to ask...

does anyone know, what is the unniversal primer for archae? i want to use it to run my PCR

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