Product Introduction:
Chloroform substitutes are mainly used to replace chloroform reagents in RNA extraction processes related to Trizol and other RNA extraction reagents. This product has low toxicity and can be used as a substitute for chloroform. The usage method is the same as chloroform; It will not have any adverse effects on the quality or quantity of isolated RNA.
Precautions：
1. If the usage is small each time, it can be packaged appropriately before use to avoid contamination.
2. For your safety and health, please wear lab clothes and disposable gloves when operating.
Instructions for Use：
1. After sample pretreatment, add 1mL of TRIzol reagent, lyse the sample, and incubate at room temperature for 5 minutes.
2. Add 0.1mL chloroform substitute, shake well, and incubate for 3 minutes.
3. Centrifuge the sample at 12000xg for 15 minutes at room temperature of 4C.
4. Take about 500uL of the upper water sample layer and transfer it to a new sample tube. After centrifugation, the mixture is divided into three layers: the lower organic layer, the middle layer, and the upper colorless water sample layer. RNA exists in the upper aqueous layer.
5. Add isopropanol of equal sample volume, invert and mix well, and incubate at room temperature for 10 minutes.
6. Centrifuge at 12000xg for 10 minutes at room temperature 4C and discard the supernatant.
7. Wash the precipitate with 1mL of 75% ethanol, centrifuge at 12000xg at room temperature for 3-5 minutes, and discard the supernatant. Repeat this step once.
Attention: The remaining small liquid can be briefly centrifuged and then sucked out with a gun tip, being careful not to discard the sediment.
8. Dry at room temperature for 3-5 minutes, add 20-50 μ L RNase free H2O, and dissolve the precipitate thoroughly. (Can dissolve rapidly in a 55 ° C water bath.)
Attention: Do not dry the sediment excessively to avoid difficulty in dissolution.
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