This product adopts Hygia C-to-C lysis and one-step Fast-script RT-PCR method, takes RNA as the template, uses gene specific primers, and performs reverse transcription and PCR amplification in the same system. No reagent needs to be added during the reaction, just add RNA template, target gene primers and water supplement system, which can effectively reduce the risk of pollution. This kit contains Fast-script one step RT mix, 2 × One step PCR master mix, where Fast-script one step RT mix includes Fast-script reverse transcriptase and RNase inhibitor. Fast-script reverse transcriptase is a mutant obtained through in vitro evolutionary screening. Its RNase H activity is reduced, and it has strong extension ability and thermal stability. 2 × One step PCR master mix includes DNA polymerase, reaction buffer, dNTPs, mg2+, RT-PCR reaction stabilizer, etc. DNA polymerase has calibration function, and the reaction product is flat end. The DNA polymerase has been modified and optimized to greatly improve the amplification performance and specificity. Under the carefully optimized reaction system and conditions, the amplification product of one step RT-PCR of this kit is up to 8 KB.
1. Please wear disposable gloves during operation to avoid RNase pollution.
2. Be sure to combine Fast-script one step RT mix and 2 × One step PCR master mix on ice.
3. Fast-script one step RT mix contains a high concentration of glycerol, which should be centrifuged briefly before use.
4. When using this kit for reverse transcription reaction, specific reverse transcription primers must be used, and random primer and oligo DT primer cannot be used.
5. It is suggested to use the RNA extracted by the kit as a template to amplify the target gene of more than 5KB.
Storage and Handling Conditions
Wet ice bag transportation; Store C-TO-C lysis reagents at room temperature, and store other components at -20℃ temperature, valid for 12 months. |
