Products | Genomics

Mastermix 16S - No False-Positive qPCR Results Anymore

Content
	Features
	DNA Contamination of Reagents
	Mastermix 16S Guarantees Ultra-Sensitive Detection
	Mastermix 16S is Conform to CE-98/79/EG and Validated
	Standard and Tailored Mastermix 16S
	Order Information

Features:

Bacterial DNA-free mastermix
Highly Active Taq Polymerase
Ultra-Sensitive Bacteria Detection
Precise Diagnosis

DNA Contamination of Reagents

Highly sensitive detection assays for bacteria are often compromised by DNA contaminations. In particular, current mastermix components carry traces of bacterial DNA, causing false-positive results.
Bacterial DNA contaminants are present essentially in all mastermix components¹:

DNA polymerase
PCR buffer
Primers
dNTPs
water

Severe problems arise with 16S and 23S rDNA qPCR. This has implications for clinical diagnosis and quality control of food and water.

A. Borst, A.T.A. Box, A.C. Fluit (2004) False-Positive Results and Contamination in Nucleic Acid Amplification Assays: Suggestions for a Destroy Strategy. Eur. J. Clin. Microbiol. Infect. Dis. 23:289-299 Pubmed

Mastermix 16S Guarantees Ultra-Sensitive Detection

Mastermix 16S relies on a proprietary technology that efficiently removes even traces of bacterial DNA from the mastermix without affecting the performance of the reagents while excluding false-positive results. The highly active MobiTaq 16S DNA polymerase allows amplifications of 40 cycles thus assuring high amplification activity. Both are essential for error-free and ultra-sensitive performance of detection assays.

With Mastermix 16S a 66% increase in detection sensitivity is achieved at 100% reliability.

Mastermix 16S is compatible with all commercial qPCR cyclers, including e.g. Roche, ABI and Bio-Rad.

Mastermix	DNA-free [%]	Percent activity
Mastermix 16S	100	100
Supplier A	12	120
Supplier B	100	34

comparison of MoBiTec Mastermix with other suppliers

Mastermix 16S is 66% more active than the mastermix of supplier B, while being DNA-free in contrast to supplier A. Amplification was run with a universal 16S rDNA primer pair for 40 cycles allowing detection of 30 genome equivalents of P. aeruginosa. - negative control w/o template; + positive control (240 pg S. aureus DNA); arrow: band position of the 610 bp amplification product.

Mastermix 16S is Conform to CE-98/79/EG and Validated

Mastermix 16S has been validated externally in qPCR sterility testing of blood products employing LightCycler® 2.0. It proved reliable in terms of true negative results and met the requirements of safe blood product release.

LightCycler® is a trademark of Roche

Standard and Tailored Mastermix 16S

MoBiTec offers a special service for a customised mastermix design:

Use of your specially developed primers
Mastermix design according to your specifications including certificate of analysis

Order Information

Mastermix 16S in its basic composition includes all reagents necessary to perform a PCR or qPCR run. Mastermix 16S is supplied as a double (2x) concentrated solution together with DNA-free deionized PCR-grade water. Mastermix 16S is also available with primers binding to conserved regions of 16S rDNA and with fluorescent dye for universal bacteria detection.

order #	description	amount
Mastermix 16S DNA-free
GE-PCR00100-01 GE-PCR00100-02 GE-PCR00100-03	Mastermix 16S Complete (including Primer &SYBR® Green)	100 Reactions 250 Reactions 1000 Reactions
GE-PCR00101-01 GE-PCR00101-02 GE-PCR00101-03	Mastermix 16S Primer (including Primer)	100 Reactions 250 Reactions 1000 Reactions
GE-PCR00102-01 GE-PCR00102-02 GE-PCR00102-03	Mastermix 16S Dye (including SYBR® Green)	100 Reactions 250 Reactions 1000 Reactions
Customized Mastermix DNA-free
GE-PCR00109-00	Mastermix Complete (including Primer &SYBR® Green)	inquire
	Mastermix Primer (including Primer)	inquire
MobiTaq 16S DNA-free DNA Polymerase
GE-POL00100-01 GE-POL00100-02	MobiTaq 16S DNA-free DNA Polymerase	100 units 500 units

SYBR® Green 1 is a trademark of Molecular Probes Inc.