EasyPure набор экстракта РНК

Описание и отзывы

Характеристики

EasyPure^® Plant RNA Kit

EasyPure^® Plant RNA Kit provides a fast and easy column based method to isolate RNA from plant tissue. Samples are lysed with detergent to inactive RNase. DNA is digested with DNase I. RNA is bound to silicamembrane. After washing, high quality RNA is eluted from the column. Isolated RNA is free of DNA and protein contamination, and is suitable for RT-PCR, qRT-PCR and northern blot.

Kit Contents

Sample requirement

Procedures

Before starting, add 48 ml 96-100% ethanol to WB6, mix thoroughly.

1. Materials preparation

Grind proper amount of plant tissues (refer to sample requirement) quickly in liquid nitrogen, add corresponding volume of BB6 (10 μl β-mercaptoethanol/ml BB6, freshly prepared), mix thoroughly by vortexing. Incubate for 3 minutes at room temperature.

2. Centrifuge at 12,000×g for 2-5 minutes and gently transfer the supernatant to a clean RNase-free microcentrifuge tube.

3. Add half volume of 96-100% ethanol in the supernatant and mix thoroughly (visible precipitates may form at this moment).

4. Vortex thoroughly to disperse the precipitates.

5. Transfer the sample (including any remaining precipitate) to the spin column. Centrifuge at 12,000×g for 30 seconds. Discard the flow-thorough (repeat this step if the volume of flow-through is larger than the volume of spin column until the entire sample is processed).

6. Add 500 μl CB6 to the spin column. Centrifuge at 12,000×g for 30 seconds. Discard the flow-thorough. Optional: When genomic DNA-free RNA is required, add 80 μl DNase I working solution (The working solution is mixed by 70 μl reaction buffer and 30 U DNase I) and incubate for 15 minutes at room temperature. Repeat step 6 once.

7. Add 500 μl WB6 (check to ensure you have added ethanol) to the spin column. Centrifuge at 12,000×g for 30 seconds. Discard the flow-thorough.

8. Repeat step 7 once.

9. Centrifuge at 12,000×g for 2 minutes at room temperature in order to remove remaining ethanol and then air-dry the column matrix for several minutes.

10. Place the spin column into a clean 1.5 ml RNase-free tube. Add 30-100 μl RNase-free Water into the center of spin column and incubate at room temperature for 1 minute.

11. Centrifuge at 12,000×g for 2 minutes to elute RNA.

12. Store the isolated RNA at -80^oC.

Notes

• All the centrifugation steps are carried out at room temperature.

• Ensure that β-mercaptoethanol has been added to BB6 before use.

• Ensure that 96-100% ethanol has been added to WB6 before use.

• This kit is not suitable for RNA purification from plant tissues rich in polysaccharide or polyphenol, only suitable for RNA purification from traditional plant tissues.

• We recommend using TransZol^TM Plant for RNA purification from plant tissues rich in polysaccharide or polyphenol.

CITATIONS 

Li F, et al. 2013. Glutamate Receptor Like Channel 3.3 is involved in mediating glutathione-triggered cytosolic Ca²⁺ transients, transcriptional changes and innate immunity responses in Arabidopsis. Plant Physiol. IF=6.555.  PMID: 23656893

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