1. Total RNA or mRNA can be used as RNA template，and we suggest using Biozol(BSC51M1)to isolate high quality RNA；
2. Quality of primers affect the performance of BioRT Master One Step RT-PCR Kit; Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special softwares.
3. RNase contamination should be avoided and follow the measures:

1） Ware one time gloves and respirator because of the RNases in saliva and skin;

2）  Use special instruments and consumables and handle in specific areas;

3） Consumables should be treated at 180℃ for 2 hours or 37℃ for 12 hours with 0.1% DEPC H2O followed by sterilized at 121℃ for 30min;

4. Enzyme mix should be slowly pipetted after centrifuging, and put back at -20℃ later;
5. Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit.
6. Enzyme mix should be slowly pipetted after centrifuging, and put back at -20℃ later;
7. Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit.