The Femto Fungal DNA Quantification Kit can be used to detect and quantify fungal DNA with high specificity and sensitivity. Fungal DNA can be reliably quantified in a background of non-fungal DNA such as bacterial, animal, and plant DNA, etc. This is essential for downstream applications that require accurate fungal DNA input including Next-Gen sequencing and metagenomic analysis. With the Femto Fungal DNA Quantification Kit, one can dependably quantify as little as 20 fg of fungal DNA in 1 µl purified biological liquids, fungal cultures, or environmental DNA samples.
Q1: Is there any general advice on GLP for qPCR?
Since this kit is so sensitive, avoiding possible sources of contamination is key for a successful qPCR.
– Work in a clean environment and decontaminate the benchtop and pipettes with a 10% bleach solution.
– Wipe down the areas with ethanol to remove the trace amounts of bleach and allow the area to try.
– Use filtered tips.
– Take care to ensure that pipetting is efficient and accurate. Calibrate your pipettes regularly.
– Add standards directly into the premix and pipet up and down to flush out any remaining liquid from the tip.
– Avoid pipetting onto the walls as this may skew the results.
– The SYTO9 dye present within the premix is sensitive to light, so working quickly and diligently is recommended.
Q2: Does qPCR Premix contain ROX?
Our qPCR Premixes does not contain ROX Reference Dye. Ensure that the ROX channel of your device is switched off.
Q3: Which settings can be used for signal detection?
Our qPCR Premixes contains Syto9. You can use the SYBR Green channel for signal detection.
Q4: Are the Femto kits certified for low bioburden?
No, kits are not certified.
Q5: What primers are used in E2007?
Primers are ITS1-F and ITS2-R.
Forward Primer: ITS-1F
5′- CTTGGTCATTTAGAGGAAGTAA-3′
Reverse Primer: ITS-2R
5′- GCTGCGTTCTTCATCGATGC-3′
They are based on this publication:
White, Thomas J., et al. “Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.” PCR protocols: a guide to methods and applications 18 (1990): 315-322
Q6: What is the primer target region?
The kit targets the ITS region
Q7: What is the origin of the fungal DNA Standards?
The DNA is derived from S. cerevisiae strain TMY18.