Product details

RNexpress spin kits help to purify high-quality RNA and non-toxic from small to large amounts of cells, tissues, and yeasts using Anewex mini-spin column. 

• Quick procedure delivering high-quality total RNA in minutes
• Ready-to-use RNA for high performance in any downstream application
• No phenol/ chloroform extraction, no CsCl gradients, no LiCl or ethanol precipitation

Applications

RNA isolated with Anewex technology has A260/280 ratios of 1.9-2.1 and are applicable for any downstream assays

• RNA-seq
• Quantitative, real-time RT-PCR
• End-point RT-PCR
• Northern, dot, and slot blotting
• Array analysis
• Poly A + RNA selection

Protocol

Purification of total RNA from animal cells

1. Collect the cells as the pellet. Cell pellets can be stored at -90 to -60^oC.
2. Disrupt the cells by adding 350 μL RL buffer to the cell pellet. Vortex or pipet to homogenize.
3. Add 350 μL Ethanol 70% to the homogenized lysate and mix well by pipetting.
4. Transfer up to 700 μL of the sample, including any precipitate that may have formed, to an RNexpress spin column placed in a 2 mL collection tube.
5. Close the lid gently, and centrifuge for 15s at ≥ 10000rpm. Discard the flow-through.
6. Add 500 μL RW1 buffer to the RNexpress spin column.
7. Close the lid gently, and centrifuge for 15s at ≥ 10000rpm. Discard the flow-through.
8. Add 500 μL RW2 buffer to the RNexpress spin column.
9. Close the lid gently, and centrifuge for 15s at ≥ 10000rpm. Discard the flow-through.
10. Place the RNexpress spin column in a new 2 mL collection tube. Close the lid gently, and centrifuge for 1 min at ≥ 10000rpm. 
11. Place the RNexpress spin column in a new 1.5 mL collection tube. Add appropriate DNAse/RNase-free water (20-50 μL) directly to the spin column membrane.
12. Close the lid gently, and centrifuge for 1 min at ≥ 10000rpm to elute RNA.