PCRmax Bacteria domain Instruction Manual

16S Ribosomal (16S) gene

Eubacteria

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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The term Eubacteria encompasses all species within the Bacteria domain. This group of
unicellular organisms has a wide range of morphologies, habitats and metabolisms, with
some being of high significance in animal and plant pathology.

The majority of bacteria are between 0.5-5µm in length with a lipid membrane surrounding
a cystoplasm lacking of organelles. Around the cell membrane is a cell wall made of
peptidoglycans. The structure of this cell wall varies between bacterial species and is used
to separate bacterial species into two groups: Gram-negative and Gram-positive. This
catagorisation can be made by applying a staining technique to a bacterial sample and
differentiating between the results.
On the outer bacterial surface there may be one or more of several appendages such as
fimbriae, pili or flagella that have different roles. Many bacteria have fimbriae over the cell
surface that are several µm in length and are thought to be used in attachment to surfaces
such as other cells. Some bacteria also have flagella which are rigid protein structures up
to 20µm in length and are used for motility.

Within the bacterial cell, genomes are not stored within a nucleus and are most commonly
found as a circular chromosome within the nucleoid. Bacterial genomes can range in size
from 160Kbp to 12.2Mbp depending on the species. Virulence factors and antibiotic
resistance are often encoded by DNA found in Plasmid formation in addition to the main
chromosome.
Bacterial reproduction is by binary fission, where a single bacterium forms two identical
daughter cells first by replication of the genome followed by division of the cell.

Introduction to Eubacteria

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MIN

MAX

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

The primers and probe sequences in this kit have 100% homology with a broad range of
Eubacteria 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

• Eubacteria specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Eubacteria 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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