Product Name HiPure Tissue DNA Mini Kit
Cat. No. & Specifications R431002,50Preps/kit
Introduction
The HiPure Universal miRNA Kit is designed for purification of total RNA, including miRNA and other small RNA molecules, from cultured cells and various animal and human tissues. This Kit combines phenol/guanidine-based lysis of samples and silica membrane-based purification of total RNA.Main Composition| Product | R431002 | R431003 |
| Preparation Times | 50 | 250 |
| HiPure RNA Mini Columns | 100 | 2 x 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| MagZol Reagent | 60 ml | 270 ml |
| Buffer RWC | 30 ml | 2 x 60 ml |
| Buffer RW2* | 20 ml | 50 ml |
| RNase Free Water | 10 ml | 30 ml |
HiPure Universal miRNA Kit components are guaranteed for at least 12 months from the date of purchase when stored at room temperature. MagZol Reagent are shipped at room temperature. After received, store at 2-8°C. Materials and Equipment to be Supplied by User1.Dilute Buffer RW2 with 100% ethanol and store at room temperature
2.Microcentrifuge capable of at least 12,000 × g
3.Chloroform (no isoamyl alcohol added)
A:Clogged HiPure RNA Column1.Too much starting material: In subsequent preparations, reduce the amount of starting material. It is essential to use the correct amount of starting material.
2.Inefficient disruption and/or homogenization: Disrupting and homogenizing starting materia as qiagen RNeasy Mini Kit pages 18-21. If working with tissues rich in proteins, we recommend using the HiPure Fibrous Tissue RNA Mini Kit.
B:RNA does not perform well (e.g. in RT-PCR1.Salt concentration in eluate too high: Modify the wash step by incubating the column for 5 min at room temperature after adding 500ul of Buffer RW2, then centriufge.
2.Eluate contains residual ethanol: Ensure that the wash flow-through is drained from the collection tube and that the column is then centrifuged at >12,000 x g for 1min.
C: DNA contamination in downstream expeiments
1.No DNase treatment: Perform optional on column DNase digestion using RNase-Free DNase Ste at the point individual protocols.
2.Incubation with Buffer RW1: In subsequent preparations, incubate the RNeasy spin column for 5 min at room temperature after addition of Buffer RW1 and before centrifuging.
D:Low A260/A280 value
1.Water used to dilute RNA for A260/A280 measurement: Use 10 mm Tris·Cl, pH 7.5, not RNAse-free water, to dilute the sample before measuring purity..