Lytic Enzyme Mixture
SIGMA/SAE0200 - for use with yeast and fungi, free of DNA contaminants, suitable for Microbiome research
Product Type: Chemical
PCR Amplification of 16S rDNA Using Universal Primer Set PCR amplification was conducted using 16S universal primers to amplify a 1,500 bp amplicon. Lanes M: 1 kb ladder (D3937) 1: SAE0200, spiked with genomic DNA 2: Heat inactivated SAE0200 3: Negative control with water only 4: Positive control containing only genomic DNA from Proteus mirabilis Heat inactivated Lytic Enzyme Mixture (SAE0200) is shown to be free of DNA contaminants by lack of amplified 16S amplicon in reaction that only contains sample (Lane 2), compared to successful amplification of the 16S amplicon in reaction spiked with genomic DNA (Lane 1).
| form | lyophilized powder |
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| Application: | The study of microbial communities has been revolutionized in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. Since DNA contamination during sample preparation is a major problem of these sequence-based approaches, DNA extraction reagents free of DNA contaminants are essential. Lytic Enzyme Mixture undergoes strict quality control testing to ensure the absence of detectable levels of contaminating microbial DNA using 35 cycles PCR amplification of 16S and 18S rDNA using universal primer sets. |
| Components: | The enzymes in Lytic Enzyme Mixture are: • Lyticase • Chitinase The enzymes are tested for absence of contaminating microbial DNA using 16S and 18S PCR amplification. |
| General description: | Lytic Enzyme Mixture is a mix of two enzymes: lyticase and chitinase, which have been proven useful in fungal and yeast lysis for microbiome studies. Lyticase is preferred to digest cell walls of yeast and generate spheroplasts from fungi for transformation. Chitinases have been detected in many microorganisms and in plants. In fungi, chitinases assist in morphogenesis, to break down the inherent chitin content of fungal cell walls. Plant chitinases help in resistance to fungal attack and counteracting fungal growth, by targeting those same fungal cell walls. In bacteria, bacterial chitinases assist in utilizing chitin as a carbon source and as an energy source. The mixture of the two enzymes has also been tested to ensure absence of microbial DNA. |
| Storage Temp. | −20°C |