Description
Product Description
Product Name HiPure Plasmid DNA Mini Kit
Cat. No. & Specifications P100102,100 Preps/Kit
Introduction
The HiPure Mini system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
Main Composition
| Product Number | P100102 | P100103 |
| Purification Times | 100 Preps | 250 Preps |
| RNase A | 5 mg | 10 mg |
| Buffer P1 | 30 ml | 80 ml |
| Buffer P2 | 30 ml | 80 ml |
| Buffer P3 | 40 ml | 100 ml |
| Buffer PW1 | 60 ml | 140 ml |
| Buffer PW2 | 20 ml | 50 ml |
| Elution Buffer | 15 ml | 30 ml |
| HiPure DNA Mini Columns II | 100 | 250 |
| 2 ml Collection Tubes | 100 | 250 |
Storage conditions and ValidityHiPure Plasmid DNA Kits components are guaranteed for at least one year when stored at room temperature.If any precipitates form in the buffers,warm at 37ºC to dissolve. After addition of RNase A and optional LyseBlue reagent, Buffer P1 is stable for 6 months when stored at 2-8°C. Preparation before Use1.Dilute Buffer PW2 with 100% ethanol and store at room temperature
2.Add the vial of RNase A to the bottle of Buffer P1 and store at 2-8˚C
3.Heat Elution Buffer to 70°C if plasmid DNA is >10kb
Troubleshooting Guide
A:Low DNA yields1.Buffer PW2 did not contain ethanol: Ethanol must be added to Buffer PW2 before used.
2.Poor cell lysis: Cells may not have been dispersed adequately prior to the addition of Buffer P2. Vortex to completely resuspend the cells.
3.Column matrix lost binding capacity during storage: Follow the Optional Protocol for Column Equilibration prior to transferring the cleared lysate to the Column. Add 100µL 3M NaOH to the column prior to loading the sample. Centrifuge at 13000 rpm for 30 seconds. Discard the filtrate.
B:Plasmid DNA floats out of well while loading agarose gel
1.Ethanol was not completely removed from column following wash steps, centrifuge column as instructed to dry the column before elution.
C:High molecular weight DNA contamination of product
1.Do not vortex or mix aggressively after adding Buffer P2. Overgrown cultures contain lysed cells and degraded DNA. Do not grow cell longer than 16 hours.
D:Absorbance of purified DNA does not accurately reflect quantity of the plasmid (A260/A280 ratio is high or low)
1.Plasmid DNA is contaminated with RNA: RNase A treatment is insufficient Confirm that the RNase A Solution was added to Buffer P1 prior to first use. The RNase A solution may degrade due to high temperatures (>65 °C) or prolonged storage (> 6 months at room temperature).
2.Background reading is high due to silica fine particulates: Spin the DNA sample at maximum speed for 1 minute; use the supernatant to repeat the absorbance readings.
Packaging & Shipping
Certificate
Laboratory
Workshop
Fininshed product area
Complaint
