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Plant RNA purification kit
Introduction
The EZgeneTM Plant total RNA kit provides an easy and fast method for isolating total RNA from plant tissues within 30 min. Only trace genomic DNA exists in the purified RNA, which can be eliminated by DNase I treatment when it is necessary.
Storage and Stability
DNase I (optional) should be stored at -20℃. All other components can be stored at room temperature. All kit components are guaranteed for 1 year from the date of purchasing.
Kit Contents
|
Catalog# |
R6611-01 |
R6611-02 |
|
Preps |
50 |
250 |
|
Buffer LY |
28 mL |
130 mL |
|
RNA Wash Buffer * |
24 mL |
112 mL |
|
DEPC-Treated ddH2O |
10 mL |
30 mL |
|
DNase Stop Buffer |
4.8 mL( included) |
24 mL( included) |
|
ezBind Columns |
50 |
250 |
|
Collection Tubes |
100 |
500 |
|
User Manual |
1 |
1 |
| Items 1 to 8 of 10 total |
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Plant RNA Mini kit
Cat#: R6611-01 Size: 50
Purification of total RNA from < 100 mg fresh or frozen plant tissue Learn More
$ 160.00
|
Plant RNA midi kit
Cat#: R6612-02 Size: 20
Purification of total RNA up to 500 microgram from 500 mg fresh or frozen plant tissue Learn More
$ 160.00
|
Plant RNA midi kit
Cat#: R6612-01 Size: 10
Purification of total RNA up to 500 microgram from 500 mg fresh or frozen plant tissue Learn More
$ 80.00
|
|
Plant RNA maxi kit
Cat#: R6614-02 Size: 25
Purification of total RNA up to 2 mg from < 5 g fresh or frozen plant tissue Learn More
$ 270.00
|
Plant RNA maxi kit
Cat#: R6614-01 Size: 10
Purification of total RNA up to 2 mg from < 5 g fresh or frozen plant tissue Learn More
$ 120.00
|
Plant RNA Mega kit
Cat#: R6615-02 Size: 10
Purification of total RNA from up to 10 mg from 20 g fresh or frozen plant tissue Learn More
$ 700.00
|
Plant RNA Mega kit
Cat#: R6615-01 Size: 2
Purification of total RNA from up to 10 mg from 20 g fresh or frozen plant tissue Learn More
$ 160.00
|
| Items 1 to 8 of 10 total |
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Show per page |
| View as: Grid List | Sort by: Best Value| Name| Price |
Before Starting
Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each steps.
Important
- R Add 1% volume of b-mercaptoethanol to Buffer LY before use and store at 4℃.
- R Add 36 mL (R6611-01) or 168 mL (R6611-02) 100% ethanol to RNA Wash Buffer before use.
- R All centrifugation steps should be carried out at room temperature.
- R Add 7.2 mL (R6611-01) or 36 mL (R6611-02) 100% ethanol to DNase Stop Buffer before use.
Materials supplied by users
- R Tabletop microcentrifuge and 1.5 mL sterile tubes.
- R Vacuum manifold if use vacuum protocol.
- R 100% ethanol
- R Optional: DNase I, DNase Buffer
Note: Perform all steps including centrifugation at room temperature
Protocol For Extracting Total RNA From Plant Tissue
- 1. Weigh 30-100 mg plant tissue in a 2 mL tube. Freeze the plant tissue in liquid nitrogen and grind using a rotor starter.
- 2. Transfer 500 µL Buffer LY to the tube containing the plant tissue immediately. Grind using a rotor starter again.
Ensure that b-mercaptoethanol has been added before use.
- 3. Centrifuge the lysate for 10 min at 14,000 rpm at room temperature and transfer the cleared lysate to a clean 1.5 mL tube.
- 4. Add 0.5 volume 100% ethanol to the lysate (for example: 250 µL 100% ethanol for 500 µl lysate).
- 5. Transfer the solution into the binding column and centrifuge at 14,000 rpm for 1 min. Discard the collection tube with the flow-through and put the column back to a new collection tube.
- 6. Add 400 µL RNA Wash Buffer to the column and centrifuge at 14,000 rpm for 1 min. Discard the flow-through.
Ensure that ethanol has been added to RNA Wash Buffer before use.
- 7. Optional: Add 50 µL DNase I (5U，RNase-free) solution onto the middle of the column and incubate at room temperature for 15 min. Add 200 µL DNase Stop Buffer onto the column and centrifuge at 14,000 rpm for 1 min. Discard the flow-through. Add 300 µL RNA Wash Buffer to the column and centrifuge at 14,000 rpm for 1 min. Discard the flow-through.
- 8. Add another 400 µL RNA Wash Buffer to the column and centrifuge at 14,000 rpm for 30 s. Discard the flow-through.
- 9. Centrifuge at 14,000 rpm for 1 min. Discard the flow-through.
- 10. Centrifuge at 14,000 rpm for another 1 min.
It is critical to remove residual ethanol for optimal.
- 11. Place the column to a RNase-free 1.5 mL tube, add 50-100 µL DEPC-treated water to the column and centrifuge at 14,000 rpm for 2 min. The RNA is in the flow-through liquid. Store the RNA solution at -20℃.
Note: It is highly recommended that RNA quality be determined before downstream applications. The quality of RNA can be assessed by denatured agarose gel electrophoresis with the ethidium bromide staining. Several sharp bands should appear on the gel including 28S and 18S ribosomal RNA bands as well as certain populations of mRNA and bands. If these bands smear towards lower molecular weight RNAs, then the RNA has undergone major degradation during preparation, handling or storage, RNA molecule less than 200 bases in length do not efficiently bind to the RNA column. An A260/A280 ratio of 1.8-2.0 corresponds to 90-100% pure nucleic acid.
Trouble Shooting Guide
|
Problem |
Possible reason |
Suggested Improvement |
|
Low A260/A280 ratios |
Protein contamination |
Do a Phenol: Chloroform extraction. Loss of total RNA (up to 40%) should be expected. |
|
Low A260/A280 ratios |
Guanidine Thiocyanate contamination |
Add 2.5 volumes of ethanol and 0.1M NaCl (final concentration) to precipitate RNA. Incubate for 30 min at -20℃. Centrifuge at 10,000 g for 15 min at 4℃. Resuspend the RNA pellet in DEPC-treated water. |
|
Low Yield |
RNA in sample degraded |
Freeze samples immediately in liquid nitrogen and store at -70℃ after collect it. |
|
Low Yield |
The binding capacity of the membrane in the spin column was exceeded |
Use of too much tissue sample exceeding the binding capacity of spin column will cause the decreasing of total RNA yield. |
|
Low Yield |
Ethanol not added to buffer |
Add ethanol to the RNA Wash Buffer and DNase Stop Solution before purification. |
|
Genomic DNA contamination |
Too much total RNA sample was used in RT-PCR. |
Reduce total RNA amount used in RT-PCR to 50-100 ng. |
|
Genomic DNA contamination |
The sample may contain too much genomic DNA. |
Reduce the amount of starting tissue in the preparation of the homogenate. Most tissues will not show a genomic DNA contamination problem at 30 mg or less per prep. Reduce cell numbers to 1-2x106 or increase buffer volume and do multiple loadings to column. |
Related EZgeneTM Products
|
Catalog # |
Product Name |
Preps |
|
R6611-01 |
Plant RNA kit |
50 |
|
R6311-02 |
Plant RNA kit |
250 |
|
R6612-01 |
Plant RNA midi kit |
10 |
|
R6612-02 |
Plant RNA midi kit |
25 |
|
R6614-01 |
Plant RNA maxi kit |
10 |
|
R6614-02 |
Plant RNA maxi kit |
25 |
|
R6615-01 |
Plant RNA mega kit |
2 |
|
R6615-01 |
Plant RNA mega kit |
10 |
|
R6815-01 |
96-well plant RNA kit |
4x96 |
|
R6815-02 |
96-well plant RNA kit |
20x96 |
|
R6411-01 |
Blood RNA mini kit |
50 |
|
R6411-02 |
Blood RNA mini kit |
250 |
|
R6412-01 |
Blood RNA midi kit |
10 |
|
R6412-02 |
Blood RNA midi kit |
25 |
|
R6414-01 |
Blood RNA maxi kit |
10 |
|
R6414-02 |
Blood RNA maxi kit |
25 |
|
R6812-01 |
96-well blood RNA kit |
4x96 |
|
R6812-02 |
96-well blood RNA kit |
20x96 |
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