Plant Genomic DNA Extraction Kit

 

Product Name	Cat.No.	Spec.
Plant Genomic DNA Extraction Kit	G3634-50T	50T

 

 

Through the use of centrifugal adsorption column technology and uniquelysis buffer system, the kit can extract genomic DNA from many kinds of simple plant tissues safely, quickly and efficiently. After complete grinding with liquid nitrogen, the plant samples were fully mixed with the lysate, which could quickly release genomic DNA, and the extraction operation was only completed within 1 hour. The kit can extract and purify 1~10 μg genomic DNA from 50~100 mg plant tissues. The extracted genomic DNA can be used in PCR amplification, Restriction endonuclease digestion, Southern hybridization, RAPD, AFLP, RFLP and other conventional molecular biology experiments.

 

 

RNase A shipped with wet ice and stored at -20°C. Other reagents were shipped and stored at room temperature; valid for up to 12 months.

 

 

Component Number	Component	G3634-50T
G3634-1	Buffer PGL1	25 mL
G3634-2	Buffer PGL2	7 mL
G3634-3	Buffer PD	9 mL
G3634-4	Buffer PW	24 mL
G3634-5	RNase A	350 μL
G3634-6	DNA Spin Columns	50个
G3634-7	Collection tubes	50个
G3634-8	Buffer TE	15 mL
Manual		One copy

 

 

1. The cryopreserved plant samples should avoid repeated freezing and thawing, otherwise the quality and yield of extracted DNA will be reduced.

2. If Buffer PGL1 precipitates,please heat it at 65°C before use until the precipitation disappears, mix well and use it.

3. Please add anhydrous ethanol (seethebottle) to Buffer PD and Buffer PW before use.

 

 

1. Lysis of plant tissues:

a. Quickly transfer 50~100 mg fresh or cryopreserved plant tissue to a 2.0 mL RNase-Free grinding tube containing 2~3 3 mm beads (recommended G0203-150G) and pre-cooled with liquid nitrogen (recommended HT-200-M). Place the grinding tube on the grinder (recommended KZ-5F-3D) (quickly pre-cool the adapter in liquid nitrogen before use) until it is powdered (if it is not completely ground to powder, it will affect the yield and quality of DNA). Then add 400 μL Buffer PGL1 and 6 μL RNase A to the grinding tube, use a pipette to blow until there is no obvious precipitation in the lysate, stand at room temperature for 10 min.

b. Quickly transfer 50~100 mg fresh or cryopreserved plant tissue to a 1.5 mL RNase-Free centrifuge tube. Add liquid nitrogen and grind the tissue with a pestle, adding liquid nitrogen continuously until the sample is ground into powder (if it is not completely ground to powder, it will affect the yield and quality of DNA). Then add 400 μL Buffer PGL1 and 6 μL RNase A to centrifuge tube, use a liquid pipette to blow until there is no obvious precipitation in the lysate, and stand at room temperature for 10 min.

c. Quickly transfer 50~100 mg fresh or cryopreserved plant tissue to a mortar pre-cooled with liquid nitrogen. Add liquid nitrogen and grind the tissue with a pestle, adding liquid nitrogen continuously until it is ground into powder (if it is not completely ground to powder, it will affect the yield and quality of DNA). The powdered sample was added to a 1.5 mL RNase-free centrifuge tube containing 400 μL Buffer PGL1 and 6 μL RNase A, use a pipette to blow until there is no obvious precipitation in the lysate and stand at room temperature for 10 min.

2. Add 130 μL Buffer PGL2 to the centrifuge tube, mix well with apipette, and place it on the ice for 5 min.

3. Centrifuge at12,000 rpm for 5 min, transfer the supernatant to a new centrifuge tube.

4. Add Buffer PD of the same volume as the supernatant to the centrifuge tube and mix it upside down.

5. Place DNA Spin Column on Collection Tube, and transfer the mixture to DNA Spin Column. Each addition does not exceed 600 μL, if exceeds, it can be added in batches.

6. Centrifugeat12,000 rpm for 30 s and discard the filtrate. Put the DNA Spin Column back into the Collection Tube.

7. Add 600 μL Buffer PW to DNA Spin Column (please add Buffer PW along the pipe wall to help flush the residual salt on the pipe wall), and centrifuge at 12,000 rpm for30 s, then discard the filtrate.

8. Repeat step 7.

9. Put DNA Spin Column into Collection Tube, centrifuge at12,000 rpmfor 2 min.

10. Transfer Spin Column to a new Nuclease-free 1.5 mL centrifuge tube. stand at room temperature for 3~5 min, so that the residual ethanol of Spin Column can be evaporate completely.

11. Add50~100 μL Buffer TE orNuclease-free Water to the centrifuge tube, stand at room temperature for5 min (heating Buffer TE or Nuclease-free Water to 65°C is beneficial to improve elution efficiency).

12. Centrifuge at 12,000 rpm for 2 min to collectDNA. To get a higher concentration ofDNA, you can also add the first eluent back toDNA Spin Column, then stand at room temperature for 5 min and centrifuge for 2 min to collectDNA again.

 

 

1. Try to use fresh plant tissue to ensure the yield and integrity of genomic DNA.

2. For long-term preservation, the extracted DNA should be eluted with Buffer TE and stored at -80°C.

3. The obtained genomic DNA should avoid repeated freezing and thawing.

 

 

The genomic DNA yield extracted from a variety of plant samples by this kit is shown in the table below. Genomic DNA yield is related to plant species, organs and growth status. The recommended dosage of samples is shown in the table below.

Sample	Dosage	DNA yield
Oryza sativaL leaf	50 mg	5~10 μg
Brassica napusL leaf	50 mg	4~8 μg
Arabidopsis thaliana leaf	80 mg	0.5~1 μg
Nicotiana tabacum leaf	50 mg	2~4 μg
Lycopersicon esculentum leaf	50 mg	2~5 μg
Allium tuberosum Rottler ex Sprengle leaf	50 mg	3~6 μg
ApiumgraveolensL leaf	50 mg	2~4 μg
Epipremnum aureum leaf	50 mg	0.5~1 μg
Populus alba leaf	50 mg	1~2 μg
Brassica rapa var. glabra Regel leaf	50 mg	1~2 μg

 

For Research Use Only!