Plant Genomic DNA Kit

Plant Genomic
DNA Kit
For purification of genomic DNA from
plant cells
www.tiangen.com/en
This product is for scientific research use only. Do not use in
medicine, clinical treatment, food or cosmetics.
DP210831
Plant Genomic DNA Kit

(Spin Column)
Cat. no. 4992201/4992202
Kit Contents
4992201 4992202
Contents
50 preps 200 preps
Buffer GP1 40 ml 160 ml
Buffer GP2 40 ml 160 ml
Buffer GD 13 ml 52 ml
Buffer PW 15 ml 50 ml
Buffer TE 15 ml 60 ml
Spin Columns CB3 50 200
Collection Tubes 2 ml 50 200
Handbook 1 1
Storage
Plant Genomic DNA Kit could be stored dry at room temperature (15-30°C)
for up to 15 months without showing any reduction in performance and
quality. If any precipitate forms in the buffers, it should be dissolved by
warming the buffers at 37°C for 10 min before use.
Plant Genomic DNA Kit Handbook

1
Introduction
The Plant Genomic DNA Kit provides a fast, simple, and cost-effective
genomic DNA extraction method for routine molecular biology laboratory
applications. The Kit uses silica membrane technology to eliminate the
cumbersome steps associated with loose resins or slurries. The kit is ready
for use and can purify the genomic DNA from a wide variety of plant
species and tissues, and the whole process is completed in less than 1
hour.
Purified DNA is suitable for PCR, restriction endonuclease digestion and
Southern Hybridization.
Yield
Sample Wet Weight DNA Yield
Plant tissues 100 mg 3-30 µg
Note: DNA yields vary with different plant samples.
Important Notes
1. Repeated freezing and thawing should be avoided; otherwise it would
reduce the DNA size and quantity.
2. If a precipitate formed in Buffer GP1 or Buffer GP2, warm buffer to 37°C
until the precipitate has fully dissolved.
Protocol
Ensure that Ethanol (96-100%) has been added into Buffer GD and Buffer
PW before use.
1. Place 100 mg wet weight plant tissue or 30 mg lyophilized plant tissue
and grind the samples thoroughly in liquid nitrogen.
2. Add 700 µl 65°C pre-heated GP1 (β-Mercaptoethanol (β-ME) should
be added to Buffer GP1 before use. The final concentration of β-ME is
0.1%) to the powdered plant tissue, Vortex for 10-20 sec to mix, make
sure to disperse all clumps and then Incubate for 20 min at 65°C, mix by
inverting the tube for several times.
3. Add 700 µl chloroform, mix by inverting the tube for several times,
centrifuge for 5 min at 12,000 rpm (~13,400 × g).
Note: If the plant sample contains polyphenol or starch, 1:1 of Phenol/
Chloroform could be used before step 3 to extract plant genomic DNA.

2
4. Pipet the supernatant water phase to a new tube, add 700 µl Buffer
GP2, mix by inverting the tube for several times.
5. Pipet all of the mixture from step 4 into the Spin Column CB3 (place
the Spin Column CB3 in the Collection Tube first). Close the CB3 lid and
centrifuge for 30 s at 12,000 rpm (~13,400 × g). Discard the filtrate and
place the Spin Column CB3 into the Collection Tube.
If the sample volume exceeds 700 µl, centrifuge successive aliquots in
the same column.
6. Add 500 μl Buffer GD (Ensure that ethanol (96-100%) is added to
Buffer GD before use), centrifuge at 12000 rpm (~13,400 × g) for 30 s
then discard the filtrate and place the Spin Column CB3 back into the
Collection Tube.
7. Add 600 μl PW (Ensure that ethanol (96-100%) is added to Buffer PW
before use) to the Spin Column CB3, and centrifuge for 30 s at 12,000
rpm (~13,400 × g), discard the flow-through, place the Spin Column CB3
back into the Collection Tube.
8. Repeat step 7.
9. Place the Spin Column CB3 in the Collection Tube, centrifuge for 2 min
at 12,000 rpm (~13,400 × g), discard the flow-through. Open lid of
CB3 and stay at room temperature for a while to dry the membrane
completely.
Note: Residual ethanol from Buffer PW may inhibit subsequent
enzymatic reactions.
10.Place Spin Column CB3 to a new centrifuge tube, and add 50-200 μl
Buffer TE directly onto the CB3 membrane, incubate for 2-5 min at room
temperature (15-30°C), and then centrifuge for 2 min at 12,000 rpm
(~13,400 × g) to elute.
Note: Elution with small volumes (<50 μl) will reduce the efficiency
of the elution and the DNA yield. The pH value of elution buffer will
have some influence in eluting; Buffer TE or distilled water (pH 7.0-
8.5) is suggested to elute DNA. For long-term storage of DNA, eluting
in Buffer TE and storing at -20°C is recommended, since DNA stored
in water is subject to acid hydrolysis. To improve the yield of genomic
DNA obtained, the elution buffer could be added to Spin Column CB3
again, stand for 2 min at room temperature (15-30°C), then centrifuge
at 12,000 rpm (~13,400 × g) for 2 min to elute.

3

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Plant Genomic

Plant Genomic DNA Kit

Cat. no. 4992201/4992202

Plant Genomic DNA Kit Handbook

Plant Genomic DNA Kit Handbook

Plant Genomic DNA Kit Handbook

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