Buffer PLY, Wash Buffer, DEPC-Treated ddH2O, ezBind Columns, Collection Tubes, User Menu
Introduction
The EZgeneTM Viral DNA/RNA kit provides an easy and reliable method for isolating total viral DNA/RNA from plasma, serum, whole blood, urine and cell culture supernatant. This procedure has been tested for isolating nucleic acids from Hepatitis A, Hepatitis C and HIV. The isolated DNA/RNA can be used for PCR, RT-PCR and other downstream applications.
Storage and Stability
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# | VR6511-00 | VR6511-01 | VR6511-02 |
| Preps | 4 | 50 | 250 |
| Buffer PLY | 2.4 mL | 28 mL | 130 mL |
| Wash Buffer * | 2 mL | 15 mL | 3 x 24 mL |
| DEPC-Treated ddH2O | 500 µL | 10 mL | 50 mL |
| ezBind Columns | 4 | 50 | 250 |
| Collection Tubes | 8 | 100 | 500 |
| User Menu | 1 | 1 | 1 |
*Add 8 mL (VR6511-00) or 60 mL (VR6511-01) or 96 mL (VR6511-02) 100% ethanol into each Wash Buffer bottle before use.
Before Starting
Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each step.
Important
- Add 1% volume of β-mercaptoethanol to Buffer PLY before use and store at 4 oC.
- Add 8 mL (VR6511-00) or 60 mL (VR6511-01) or 96 mL (VR6511-02) 100% ethanol into each Wash Buffer bottle before use.
Materials supplied by users
- Tabletop microcentrifuge
- 1.5 mL sterile tubes
- 100% ethanol
Note: Perform all steps including centrifugation at room temperature
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°C. Centrifuge at 13,000 rpm for 15 min at 4°C. Resuspend the RNA pellet in DEPC-treated water. |
| Low Yield | RNA in sample degraded
|
Freeze samples immediately in liquid nitrogen and store at -70°C after collect it. |
| Low Yield | The binding capacity of the membrane in the spin column was exceeded | Use of too much 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 Wash Buffer. |
| 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. |
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