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CTAB Plant gDNA Extraction Kit(SpinColumn)

Packing Specification：

Product Introduction:
This rapid plant genomic DNA extraction kit adopts an improved CTAB method, suitable for extracting genomic DNA from plants in approximately 60 minutes. The unique lysis buffer can rapidly lyse cells. Combined with chloroform extraction, isopropanol centrifugal precipitation, and multiple column washing steps, high-purity, high-quality genomic DNA free from impurities such as proteins and cellular metabolites can be extracted. Typically, the yield from 100 mg of fresh tissue ranges from 3~25 μg. It can be used for various molecular biology experiments such as enzyme digestion, PCR, Southern Blot, library construction, and molecular marking.

Product Features:
1.No contamination: Contains RNA digestive enzymes to effectively remove RNA from genomic DNA.
2.Special applicability: Suitable for extracting plant tissues rich in phenols and polysaccharides.
3.Easy operation: Simple workflow, with the entire extraction process taking only about 60 minutes.
4.Spin column: Equipped with specific adsorption columns for extraction through washing-centrifugation-elution steps.
5.High quality: Effectively ensures DNA integrity, with high recovery efficiency and purity. Free from RNA and other impurities, the OD260/OD280 ratio can reach 1.7~1.9, suitable for various molecular biology experiments.

Application Scope:
Suitable for the rapid extraction of genomic DNA from various plant tissues, leaves, roots, stems, and whole plants.

Operation Steps:
Tips:
Before first use, add the specified amount of anhydrous ethanol to Wash Buffer WB, mix thoroughly, and mark the box to indicate ethanol has been added to avoid repeated addition!
Preheat the required amount of Lysis Buffer PL at 65℃ before the experiment. Add β-mercaptoethanol to a final concentration of 2% before use (e.g., add 20 μL of β-mercaptoethanol per 1 mL of Lysis Buffer PL).
Preheat Elution Buffer EB in a 65℃~70℃ water bath.

1.Take an appropriate amount of plant tissue (100 mg of fresh tissue or 30 mg of dry tissue) and grind it thoroughly into fine powder in a mortar with liquid nitrogen. Note: Due to the high diversity of plants and varying DNA content in different growth stages and tissues of the same plant, select the appropriate amount of plant material based on specific experimental conditions.

2.Transfer the powder to a 1.5 mL centrifuge tube. Add 600 μL of preheated Lysis Buffer PL (65℃) (ensure β-mercaptoethanol has been added) and mix thoroughly for lysis. Gentle homogenization for 10 seconds can be performed if necessary.

3.Incubate in a 65℃ water bath for 20~60 minutes, inverting the centrifuge tube 2~3 times during incubation to mix thoroughly. If RNA residue is high, add 6 μL of RNase (20 mg/mL) before water bath incubation.

4.Add 700 μL of chloroform or chloroform/isoamyl alcohol (mixed at a volume ratio of 24:1), invert to mix thoroughly for a few minutes (or vortex to mix), then centrifuge at 13,000 rpm for 5 minutes. For plant tissues rich in polysaccharides and polyphenols, extract once with an equal volume of phenol/chloroform (1:1) before Step 4.

5.Carefully aspirate the supernatant into a new 1.5 mL centrifuge tube, taking care not to aspirate the interface material. If the supernatant is turbid, repeat Step 4 until a clear supernatant is obtained.

6.Add 1.5 volumes of Binding Buffer PQ (ensure anhydrous ethanol has been added) to the supernatant and immediately vortex to mix thoroughly.

7.Load the mixed solution (700 μL each time) into a Spin AC column sleeved in a collection tube. Centrifuge at 13,000 rpm for 30 seconds and discard the waste liquid.

8.Add 500 μL of Inhibitor Removal Buffer IR, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

9.Add 600 μL of Wash Buffer WB (ensure anhydrous ethanol has been added), centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

10.Repeat Step 9 once.

11.Place the Spin AC column back into the collection tube and centrifuge at 13,000 rpm for 2 minutes to thoroughly remove residual Wash Buffer (residual ethanol may inhibit downstream reactions).

Remove the Spin AC column and place it into a clean centrifuge tube. Add 100 μL of preheated Elution Buffer EB to the center of the adsorption membrane. Incubate at room temperature for 3-5 minutes, then centrifuge at 12,000 rpm for 1 minute. Transfer the eluted solution back into the spin column, incubate at room temperature for 2 minutes, and centrifuge at 12,000 rpm for 1 minute.