Total RNA Extraction Kit - Tissue
For total RNA Extraction Kit - Tissue
Size: 10 tests
Size: 10 tests
型號 :
RT10
Total
RNA Extraction Kit – Tissue is designed by patented technology for purifying total RNA from varied animal
tissues. This method uses detergents and a chaotropic salt to lyse cells and
inactivate RNase. RNA in the chaotropic salt solution binds to the glass fiber
matrix (NAB Filter) of 
NAB
Nanosep® Device. Following washing off contaminants, purified RNA is eluted by
RNase-free water. ssRNA and dsRNA can be efficiently purified. Purified RNA is
ready for RT-PCR, northern blotting, primer extension and cDNA library
construction.
|
Preparation Before Assay |
▓ Add 10 μl ß-ME to 1 mL of RL Buffer. Note: Prepared RL Buffer can be stored at room temperature for up to 1 month. |
|
Sample Preparation |
① Cut and place 10 mg of fresh or frozen animal tissue in a RNase-free microcentrifuge tube. ② Add 400 μl of RL Buffer (ß-ME added) into the tube and sufficiently grind the tissue. Optional: Shear the tissue by passing lysate though a 20-G needle syringe for 10 times. Note: If insolubles remain after incubation, centrifuge for 2 minutes at 13,000 rpm (10,000 x g) and transfer the clarified supernatant to the new microcentrifuge tube. |
|
Assay Procedures |
① Add 400 μl of RB Buffer to the sample and mix by pipetting immediately for 10 seconds. ② Transfer 450 μl of the mixture to Nanosep®. ③ Centrifuge at 13,000 rpm (10,000 x g) for 3 minutes. ④ Remove the retentate cup of Nanosep®, discard the filtrate from the filtrate tube, and then place the retentate cup back into the filtrate tube of Nanosep®.
⑤ Transfer the remaining mixture to Nanosep®. ⑥ Repeat ③ and ④. Optional: Perform DNA Elimination Procedures between ⑥and ⑦if needed. ⑦ Add 450 μl of RW1 Buffer to Nanosep®. ⑧ Centrifuge at 13,000 rpm (10,000 x g) for 1 minute. ⑨ Remove the retentate cup of Nanosep®, discard the filtrate from the filtrate tube, and then place the retentate cup back into the filtrate tube of Nanosep®. ⑩ Add 450 μl of RW2 Buffer to Nanosep®. ⑪ Centrifuge at 13,000 rpm (10,000 x g) for 1 minute. ⑫ Remove the retentate cup of Nanosep®, discard the filtrate from the filtrate tube, and then place the retentate cup back into the filtrate tube of Nanosep®. ⑬ Repeat ⑩, ⑪ and ⑫. ⑭ Centrifuge at 13,000 rpm (10,000 x g) for 3 minutes to dry NAB Filter. ⑮ Remove the retentate cup of Nanosep®, and transfer it into the new filtrate tube. ⑯ Add 50 μl of RNase-Free Water into the CENTER of the retentate cup. ⑰ Let the device stand for at least 2 minutes so NAB Filter can be soaked completely. ⑱ Centrifuge at 13,000 rpm (10,000 x g) for 2 minutes to elute the purified RNA. |