Fast Cell RNA Extraction Kit (Chloroform-free / β-Mercaptoethanol-free)

1. Before first use，add absolute ethanol to the Wash Solution RW as instructed on the bottle label.2. The lysis buffer is irritating to the skin. Wear latex gloves and a mask during operation to avoid contact with skin，eyes and clothing. In case of contact，rinse immediately with clean water.3. Use RNase-free reagents and consumables to prevent cross-contamination. Keep the laboratory clean to avoid RNase contamination.4. Perform all centrifugation steps at room temperature using a benchtop centrifuge with a maximum speed of 13，000 rpm.5. The cell number should be controlled between 5×10??and 5×10?. RNA content varies greatly among different cell types. For reference，1×10??HeLa cells yield 15 μg RNA，while 1×10??COS-7 cells yield 35 μg RNA. If the RNA content of the sample cells is unknown，perform a gradient test to determine the optimal cell quantity.6. After mixing cells with lysis buffer，the lysate can be stored at -20 °C if the subsequent procedures cannot be carried out immediately. Storage at -20 °C for 16 hours will reduce RNA yield by approximately 20% without obvious RNA degradation. Do not leave the lysate at room temperature for a long time. Tests show that RNA degrades significantly after 1 hour at 37 °C. For large batches of samples，process them in groups or complete the follow-up steps within 15 minutes.7. Trace DNA residue: The proprietary lysis buffer of this kit removes most genomic DNA，so additional DNase digestion is not required. The extracted RNA can be directly used for RT-PCR and qRT-PCR. If excessive DNA residue occurs or highly stringent downstream experiments are required，perform DNase I digestion after Step 5. This kit contains a proprietary lysis buffer that efficiently lyses cells and inactivates cellular RNases. It greatly shortens extraction time，reduces the risk of RNA degradation and improves RNA integrity. Total RNA is selectively bound to the silica membrane of the spin column under high-salt conditions. Residual proteins，cellular metabolites and other impurities are then removed via washing steps. Finally，pure RNA is eluted from the silica membrane using RNase-free water with low salt concentration. Product Advantages:1. Non-toxic & eco-friendly: No toxic reagents such as phenol，chloroform or β-mercaptoethanol are involved，and ethanol precipitation is not required. 2. Fast & simple workflow: Fewer operating steps. RNA extraction for a single sample can be completed within 15 minutes. 3. High-quality RNA: The extracted total RNA maintains excellent integrity. The typical OD260/OD280 ratio ranges from 2. 1~2. 2 with nearly no DNA contamination. The product is fully applicable to downstream experiments including RT-PCR and Northern blot.