FAQ DNA-free PCR Reagents
Can I use Molzym's PCR assays for quantitative PCR (qPCR) analysis?
Products: Mastermix 16S Complete, Mastermix 18S Complete, PCR assays incl. in SepsiTest™-UMD, Micro-Dx™ and UMD-SelectNA™ No. Due to the broad-range primers (16S/18S) used, potential primer dimer formation and unspecific annealling can occur, so we ...
Does Molzym manufacture customized mastermixes?
Customized Mastermix Yes. Please find further information here and send inquiries on customized manufactured master mixes to info@molzym.com
The melt curve analysis shows a high primer dimer formation: why and what can I do?
Primer dimer Primer dimer formations are non-specific products and can be minimized by constant cooling during PCR setup. It is completely normal that in negative samples, for example, the negative control (NC), the primer dimer can be higher ...
How to interpret the PCR results?
Amplicons can either be analyzed by conventional gel electrophoresis with bands displaying at the size of the amplicons (460bp for the Mastermix 16S Complete and 310bp for the Mastermix 18S Complete). Alternatively, a real-time PCR cycler can be ...
How can bacteria and fungi be identified when detected by 16S or 18S rDNA PCR?
Identification from positive amplicons The direct way is (Sanger) sequence analysis. Amplicons are purified in a simple procedure, using a commercial PCR purification kit (e.g. QIAQuick PCR Purification Kit, Qiagen, Germany). The purified amplicon is ...
Which reagents can bear contamination by microbial DNA?
In principle, all! In particular, in our experience, the Taq DNA polymerase and primers are mostly prone to DNA contamination due to the manufacturing processes. Molzym has invested great efforts to develop a purification process for the elimination ...
What is the sensitivity of detection?
The analytical sensitivity of the assay Mastermix 16S Complete is strain-dependent, please find below some examples: Strain CFU/ml Escherichia coli 40 Klebsiella pneumoniae 50 Staphylococcus aureus 20 Streptococcus pneumoniae 50 Enterococcus faecalis ...
Why is broad-range 16S rDNA PCR a good option for the detection of bacteria?
Broad-range 16S rDNA PCR is a favorable way of detecting bacteria in samples due to two reasons. First, the use of highly conserved regions in the 16S rRNA gene allows the amplification of sequences from any bacterial strain. Mastermix 16S Complete ...
What are the differences between the DNA-free master mixes and assays?
The assays Mastermix 16S Complete and Mastermix 18S Complete include already primers allowing for the ultra-sensitive detection of bacterial DNA (V3/V4 variable region of the 16S rRNA gene) and fungal DNA (V8/V9 variable region of the 18S rRNA gene) ...
Is MolTaq 16S/18S a hot-start Taq polymerase?
Molzym’s hot-start Taq DNA Polymerase is called HotMolTaq 16S/18S (order number P-080-0100/P-080-0500) and is aptamer-stabilized for setup at room temperature. MolTaq 16S/18S (order number P-019-0100/P-019-0500) on the other hand is not ...
Can MolTaq 16S/18S and master mixes be used for Real-Time PCR?
MolTaq 16S/18S & real-time PCR Yes. All DNA-free PCR products of Molzym are fully compatible and optimized for Real-Time PCR, using fluorescent dye or probe detection systems.
What does “DNA-free” mean?
“DNA-free” products by Molzym are defined as reagents and plastics being negative for bacterial and fungal DNA for up to 40 PCR cycles using broad-range bacterial and fungal PCR assays (Mastermix 16S Complete and Mastermix 18S Complete). Therefore, a ...