Fungi/Yeast Genomic DNA Isolation Kit

Supplier
Norgen Biotek
Cat. no.
1027300
Kit size
50
Links
Norgen Biotek Protocol Information Sheet

Features

• Isolate DNA from a wide range of samples - Genomic DNA can be isolated from viable yeast cells, fungal spores or mycelium and Gram-positive bacteria.
• No phenol :chloroform extractions - DNA is isolated without the use of harmful chemicals such as phenol or chloroform.
• Rapid and simple processing - Rapid bead-beating and spin-column format allows for the processing of multiple samples in 45 minutes.
• late high quality DNA - Purified DNA is of the highest quality and can be used in a number of downstream applications including PCR, Restriction Fragment Length Polymorphism (RFLP) and Amplified Fragment Length Polymorphism (AFLP) .

Norgen’s Fungi/Yeast Genomic DNA Isolation Kit is designed for the rapid preparation of genomic DNA from viable yeast cells, fungal spores or mycelium and Gram-positive bacteria. Genomic DNA is efficiently extracted from the cells by a combination of heat treatment, detergents and the use of provided Bead Tubes. The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with downstream applications such as PCR and Southern Blot analysis. Typical yields of genomic DNA will vary depending on the cell density of the yeast or fungal culture and species. The option of an additional lyticase treatment is also provided in order to allow for improved DNA yields for certain fungal and yeast species. Preparation time for a single sample is less then 30 minutes, and each kit contains sufficient materials for 50 preparations.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. The process involves first adding Lysis Solution to the fungi or yeast sample. At this point, an optional RNase treatment can be performed if RNA-free genomic DNA is required. Next, the mixture is placed into a provided Bead Tube and the tube is vortexed for 5 minutes in order to efficiently and rapidly homogenize the sample and extract the DNA. The sample is then centrifuged, and the supernatant is transferred to a DNAse-free microcentrifuge tube. Ethanol is added to the lysate, and it is then loaded onto one of the provided spin columns containing Norgen’s proprietary resin. Norgen’s resin binds nucleic acids in a manner that depends on ionic concentrations, thus only the DNA will bind to the column while the proteins are removed in the flowthrough or retained on top of the resin. The bound DNA is then washed twice using the provided Wash Solution, and the purified DNA is eluted using the Elution Buffer. The purified DNA can be used in sensitive downstream applications including PCR and Southern blotting.

Column Binding Capacity	50 µg
Maximum Column Loading Volume	600 µL
Maximum Amount of Starting Material:	Fungi (wet weight): 50 mg	Yeast or bacterial culture: 0.5-1 mL
Time to Complete 10 Purifications	30 minutes

Isolate DNA from a Wide Range of Samples

DNA was isolated from 50 mg (wet weight) samples of Pichia sp. (Lane 1), Aspergillus niger (Lane 2), Cladosporium cladasporioides (Lane 3), Botrytis cinerea (Lane 4) and Mucor racemosus (Lane 5) using Norgen’s Fungi/Yeast Genomic DNA Isolation Kit, and 5 µL aliquots were run on a 1X TAE 1% agarose gel. As it can be seen, high quality DNA was isolated in all cases. The M Lane contains Norgen’s HighRanger 1kb DNA Ladder.

Purified DNA can be Amplified in a qPCR Reaction

DNA was isolated from 50 mg samples of Pichia sp. , Aspergillus niger , Cladosporium cladasporioides , Botrytis cinerea and Mucor racemosus using Norgen’s Fungi/Yeast Genomic DNA Isolation Kit, and 2 µL of the DNA was used in a qPCR (SYBR Green) reaction with specific fungal and yeast primers. The qPCR was successful in amplifying and detecting all the yeast and fungal DNA, indicating that the DNA is of a high quality and can be used in sensitive downstream applications.

• PCR
• Restriction Fragment Length Polymorphism (RFLP)
• Amplified Fragment Length Polymorphism (AFLP)