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Biochemical and Molecular Test

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

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Biochemical and Molecular Test

Postby sarahjohn2005 » Thu Oct 27, 2011 4:53 am

A dog's mouth is cleaner (micro biologically) than a human's mouth. How can you experimentally test this using Molecular technique ? Thanks
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Postby JackBean » Thu Oct 27, 2011 6:43 am

take little of salive and put it onto agar plate and incubate
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Postby canalon » Thu Oct 27, 2011 5:43 pm

Universal 16S amplification followed by DGGE or TGGE.
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Re: Biochemical and Molecular Test

Postby Brittney » Fri Oct 28, 2011 1:05 am

sarahjohn2005 wrote:A dog's mouth is cleaner (micro biologically) than a human's mouth. How can you experimentally test this using Molecular technique ? Thanks

um i think use a tissue to take saliva from the dog and human and test it :? good question
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Postby daniel.kurz » Fri Oct 28, 2011 5:55 am

Define clean. Clean as in sterile, clean as in clear of possible or confirmed pathogenic species, clean as in devoid of those microbes that make plaques. The frame of reference matters.
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Re: Biochemical and Molecular Test

Postby sykierj » Wed Nov 16, 2011 7:12 am

sarahjohn2005 wrote:A dog's mouth is cleaner (micro biologically) than a human's mouth. How can you experimentally test this using Molecular technique ? Thanks


hi sarahjohn2005, as you are looking for test on molecular technique i don't know whether you understand or not but what i want to share, you know it's one of the most basic techniques of molecular biology to study protein function is expression cloning. In this technique, DNA coding for a protein of interest is cloned (using PCR and/or restriction enzymes) into a plasmid (known as an expression vector). This plasmid may have special promoter elements to drive production of the protein of interest, and may also have antibiotic resistance markers to help follow the plasmid.

This plasmid can be inserted into either bacterial or animal cells. Introducing DNA into bacterial cells can be done by transformation (via uptake of naked DNA), conjugation (via cell-cell contact) or by transduction (via viral vector). Introducing DNA into eukaryotic cells, such as animal cells, by physical or chemical means is called transfection. Several different transfection techniques are available, such as calcium phosphate transfection, electroporation, microinjection and liposome transfection. DNA can also be introduced into eukaryotic cells using viruses or bacteria as carriers, the latter is sometimes called bactofection and in particular uses Agrobacterium tumefaciens. The plasmid may be integrated into the genome, resulting in a stable transfection, or may remain independent of the genome, called transient transfection.

In either case, DNA coding for a protein of interest is now inside a cell, and the protein can now be expressed. A variety of systems, such as inducible promoters and specific cell-signaling factors, are available to help express the protein of interest at high levels. Large quantities of a protein can then be extracted from the bacterial or eukaryotic cell. The protein can be tested for enzymatic activity under a variety of situations, the protein may be crystallized so its tertiary structure can be studied, or, in the pharmaceutical industry, the activity of new drugs against the protein can be studied.
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Postby JackBean » Wed Nov 16, 2011 3:04 pm

how will protein expression help to determine amount of bacteria in saliva? :roll:
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