Animal and Fungi DNA Preparation Kit solution-based | DNA Genômico Cellco

Animal and Fungi DNA Preparation Kit - solution-based

Ficha técnica

Código	Animal and Fungi DNA Preparation Kit - solution-based
Categoria	DNA Genômico - Kits baseados em soluções
Marca	Cellco

Descrição Geral

Animal and Fungi DNA Preparation Kit - solution-based | DNA Genômico - Cellco
Solution based genomic DNA purification from animal tissue and fungi

For in vitro use only!

Envio:

shipped at ambient temperature

Condições de armazenamento:

store at ambient temperature

Validade:

12 months

Descrição:

Animal and Fungi DNA Preparation Kit is designed for convenient and fast isolation of genomic DNA from animal tissue and fungi. The solution based system minimizes DNA fragmentation that may be problematic in spin-column / filtration based methods. Because phenol or chloroform is not used it is safe and does not produce any harmful waste.

Solution based genomic DNA purification kits guarantee minimal DNA fragmentation and yield DNA sized up to 150 kb.

Kit Content:

Cell Lysis Solution

Cell Resuspension Solution

Proteinase K (before use, solve in double distilled water to obtain a final concentration of 20 mg/ml) - store at -20 °C

Protein Precipitation Solution

Washing Buffer (before use, add 96-99 % Ethanol as indicated on the bottle)

DNA Hydration Solution

RNase A (before use, solve in double distilled water to obtain a final concentration of 4 mg/ml) - store at -20 °C

Additional Materials Required:

Isopropanol (2-propanol) >99 %

96-99 % Ethanol

Microtubes 1.5 ml

Preparation procedure:

Before start, provide >99 % Isopropanol (2-propanol) (not included in the kit).

For S pack (100 preps): Add 200 μl dd-water to the Proteinase K tube, 200 μl dd-water to the RNase A tube and 48 ml 96-99 % Ethanol (not included in the kit) to the Washing Buffer bottle.

For L pack (500 preps): Add 200 μl dd-water to each Proteinase K tube, 200 μl dd-water to each RNase A tube and 120 ml 96-99 % Ethanol (not included in the kit) to each Washing Buffer bottle.

Buffer	PP-208S 100 preps	PP-208L 500 preps
Cell Lysis Solution	32 ml	160 ml
Cell Resuspension Solution	32 ml	160 ml
Proteinase K (20 mg/ml)	4 mg	5x 4 mg
Protein Precipitation Solution	11 ml	55 ml
Washing Buffer	add 48 ml Ethanol (final volume 60 ml)	add 120 ml Ethanol to each bottle (final volume 150 ml each)
DNA Hydration Solution	11 ml	55 ml
RNase A (4 mg/ml)	0.8 mg	5x 0.8 mg

1a Cell Lysis for Animal Tissue:

Transfer 5-10 mg of fresh or frozen tissue to a 1.5 ml microtube.

Add 300 μl Cell Lysis Solution to the tissue.

Add 1.5 μl Proteinase K Solution to the lysate and mix by inverting several times.

Incubate at 55 °C overnight or until tissue has dissolved.

1b Cell Lysis for Fungi:

Transfer 1 ml of the cultured cells to a 1.5 ml microtube.

Harvest the cells by centrifuging at 15,000 g for 1 min and discard supernatant.

Resuspend the cell pellet in 300 μl Cell Resuspension Solution.

Add 1.5 μl Proteinase K Solution and mix by inverting several times.

Incubate at 55 °C for 60 min.

Centrifuging at 15,000 g for 1 min and discard supernatant.

Resuspend the pellet in 300 μl Cell Lysis Solution.

2 Protein Precipitation:

Add 100 μl of Protein Precipitation Solution to the cell lysate.

Mix the solution well by vortexing for 20 sec.

Centrifuge at 15,000 g for 3 min. (The precipitated protein will be a tight pellet. If the pellet is not tight, repeat mixing, incubate on ice for 10 minutes, and then centrifuge again.)

3 DNA Precipitation:

Transfer the supernatant to a clean 1.5 ml microtube containing 300 μl of Isopropanol >99 %.

Mix the sample by inverting gently 50 times.

Centrifuge at 15,000 g for 1 min. The DNA will be visible as a pellet that ranges in color form off-white to light green.

Discard the supernatant and drain tube briefly on clean absorbent paper.

Add 500 μl Washing Buffer and invert tube several times to wash the DNA pellet.

Centrifuge at 15,000 g for 1 min. Discard the ethanol carefully.

Air dry at room temperature for 10-15 min.

4 DNA Hydration:

Add 50-100 μl of DNA Hydration Solution to the dried DNA pellet.

Add 1.5 μl of RNase A to the cell lysate.

Mix the sample by inverting the tube and incubate at 37 °C for 30-60 min.

Hydrate the DNA by incubating sample at 65 °C for 60 min.

Store DNA at 4 °C. For long time storage, place sample at -20 °C or -80 °C.

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