PCRmax Oxalobacter formigenes Instruction Manual

Oxalyl-CoA decarboxylase gene

Oxalobacter
formigenes

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Oxalobacter formigenes is an oxalate-degrading anaerobic bacterium that colonizes the
large intestine of numerous vertebrates, including humans. O. formigenes and humans
share a beneficial symbiosis.

O. formigenes is a rod-shaped, gram negative obligate anaerobe. O. formigenes is a non­motile, non-spore-forming, gram negative beta-proteobacterium that colonizes the human
large intestine (colon)
O. formigenes is a strict anaerobe and is thought to be largely important in the metabolism
of oxalate in the large intestine that may be remaining from the diet or a by-product of host
metabolism.

The absence of O. formigenes is thought to be a cause of hyperoxaluria and the formation
of calcium oxalate stones in the kidney and it is thought that O. formigenes may be useful
as a probiotic agent for the treatment of these conditions.
I.E. If a person's gastrointestinal tract lacks this bacterium, and therefore lacks the primary
source for the Oxalyl-CoA decarboxylase enzyme, then the GI tract cannot degrade
calcium oxalate. Such an imbalance may facilitate the formation of Calcium Oxalate based
kidney stones

O. formigenes requires oxalate as a source of carbon and energy in order to survive. Most
anaerobes that metabolize oxalate produce carbon dioxide and formate, and the formate is
then further oxidized by dehydrogenases. O. formigenes is unique in that it produces
formate as an end product. The oxalate taken in by the cell can be decarboxylated to
regenerate formate for export, or further metabolized for the production of glycerate.

Additionally, O. formigenes has not been documented to utilize glycolytic pathways, and is
thus unable to grow on sugars

Introduction to Oxalobacter formigenes

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MIN

MAX

The PCR Max qPCR Kit for Oxalobacter formigenes (O.formigenes) genomes is designed
for the in vitro quantification of O.formigenes genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the O.formigenes genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
O.formigenes sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

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Kit Contents

• O.formigenes specific primer/probe mix (150 reactions BROWN)

FAM labelled

• O.formigenes positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

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