PCR Quantification
Standards
The quantification standard is genomic DNA extracted from low passage
and defined mycoplasma or legionella cultivated in liquid medium according
to the European Pharmacopoeia or in Hayflick´s medium.
The extraction of the DNA is done by column absorption methods and the
DNA purified by phenol/chloroform extraction and ethanol precipitation.
The concentration was determined photometrically and by real-time PCR.
The DNA concentration was adjusted by dilution with DNA stabilizing buffer.
Genomic DNA is used as a control template when DNA amplification tests
by conventional PCR are performed (e.g. as contamination control in cell
culturing). With the real-time PCR technology, this titrated DNA standard
with a defined concentration of genome copies is used as calibrator to
generate standard curves. Therefore the provided solution is diluted in
a 10x dilution series with DNA-free water and used as a PCR sample. With
the generated fluorescent data, the software provided with the real-time
PCR instrument calculates a standard curve, which can be used to determine
the DNA concentration of unknown samples.
Applications
These DNA-Standards allow for the performance of a control reaction for
PCR using real-time PCR instrumentation and to create a standard curve
for quantification experiments, i.e. with Venor®Mp
Mycoplasma pneumoniae Diagnostic Kit for real-time PCR.
The DNA standards can be used as a general control to confirm PCR system
performance using a real-time PCR instrument or for the validation of
detection limits in respect to conventional PCR tests.
Contents
1 vial (green cap) of Standard DNA, 100 µl, contains approx. 106
genomes/µl.
3 vials (white cap) with 1 ml of DNA free water.
Ordering
| DNA |
Cat.-No. |
| Legionella pneumophila DNA (DSMZ 7513 / ATCC 33152 / strain
Philadelphia-1) |
52-0101 |
| Mycoplasma orale DNA (NCTC 010130) |
52-0112 |
| Mycoplasma pneumoniae DNA (NCTC 010119) |
52-0119 |
| Acholeplasma laidlawii |
52-0116 |
|
