Q1: Why is the column clogged?
A1：
1) The amount of sample is too much. Take an appropriate amount of sampling according to the requirements recommended in the instructions.
2) If the lysate is viscous and difficult to absorb, the sample can be centrifuged at 12,000 rpm (~13,400×g) for 2 minutes before step 2, and the supernatant can be filtered before step 2;
3) The sample is rich in polysaccharides and polyphenols. For tissues rich in polysaccharides and polyphenols, it is recommended to use a special kit for extraction; 4) The centrifuge temperature is too low. The operation of this product is carried out at room temperature.
Q2：Why is the extracted RNA degraded?
A2：
1) Too much sample is used. It affects the lysing ability of the lysate, resulting in insufficient inhibition of RNase, resulting in RNA degradation. It is recommended to refer to the recommended sampling volume in the manual. If the initial sample volume is to be increased, the volume of each solution in subsequent experiments must be increased in equal proportions. For tissues with high endogenous RNase content, the sample volume should be reduced, and the amount of lysate should be increased appropriately;
2) Improper storage of samples. Repeated thawing will cause RNA degradation, try to use fresh samples or samples that have been thawed no more than twice;
3) RNase contamination was introduced during operation. Please use RNase-Free tools, reagents and consumables; 4) Degradation occurs during electrophoresis. Use RNase-Free Loading Buffer, agarose and electrophoresis buffer, etc.
Q3：Why is the RNA yield low?
A3：
1) Too much sample is used. Excessive sample will affect the cracking effect, it is recommended to take appropriate amount of samples according to the instruction manual;
2) Insufficient sample lysis. Please fully grind and crack according to the requirements of the instructions;
3) Insufficient elution. RNase-Free ddH2O needs to be directly added to the center of the membrane, and then centrifuged after standing for 2 minutes. If necessary, a second elution can be performed to increase the yield;
4) There is ethanol residue in the adsorption column. After rinsing with Buffer GRW2, it is necessary to open the cover to dry the ethanol in the adsorption column.
Q4：Why is there DNA contamination in the extracted RNA?
A4：
1) Too much sample is used. Exceeding the sample volume specified in the kit affects the lysing ability of the lysate, which may lead to genome contamination;
2) There are many secondary metabolites. Samples with high content of secondary metabolites are prone to genome contamination during RNA extraction;
3) During the operation, it is necessary to remove genomic DNA. If the DNA content is large, the DNase I digestion time can be extended or the digestion can be repeated.
