What is SuperScript IV?
SuperScript® IV Reverse Transcriptase (RT) is a proprietary MMLV mutant with superior robustness and reliability in RT reactions. SuperScript® IV RT is the top choice for all RT-PCR and qRT-PCR applications in the SuperScript® product family.
What is the difference between SuperScript III and IV?
Are there any significant changes in the SuperScript IV RT protocol compared to the SuperScript III RT protocol? The only change is that the incubation time for the reverse transcription reaction has been reduced from 50 minutes to 10 minutes. All the other parameters and steps are the same.
What is SuperScript III?
The SuperScript® III First-Strand Synthesis System for RT-PCR is optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA. SuperScript® III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability.
What is reverse transcribed into cDNA?
Reverse transcription is a technique used by researchers to generate a complementary strand of DNA (cDNA) from RNA. In this situation, researchers can isolate mRNA from the tissue and then use reverse transcription to produce cDNA.
What is RT protocol?
Reverse transcription (RT) is the process of converting RNA to cDNA using a reverse transcriptase enzyme and dNTPs. The following experiments can be used as basic RT protocols that can be modified to suit particular requirements.
What is the difference between SuperScript II and III?
SuperScript® III RT is relatively thermostable and has a standard reaction temperature of 50°C. This is 8°C higher than that of SuperScript® II and GoScript™ reverse transcriptases (42°C) and 13°C higher than that recommended for Omniscript® or Sensiscript® RTs (37°C).
What is the best reverse transcriptase?
Conclusions. Our results suggest that, of the 11 reverse transcriptase enzymes subjected to our investigation, the Accuscript (ACC) and Superscript III (SS3) were the best performing enzymes in terms of reproducibility and sensitivity for low copy RNA levels.
Why do we need to reverse transcribe the mRNA to cDNA?
The synthesis of DNA from an RNA template, via reverse transcription, produces complementary DNA (cDNA). This combination of reverse transcription and PCR (RT-PCR) allows the detection of low abundance RNAs in a sample, and production of the corresponding cDNA, thereby facilitating the cloning of low copy genes.
Which is the best reverse transcriptase for cDNA synthesis?
The Invitrogen SuperScript line of reverse transcriptases (RTs) represents the most widely used and highly referenced RTs for cDNA synthesis in many applications. Invitrogen Superscript IV RT, the most recently introduced enzyme, was engineered to offer superior cDNA synthesis performance with even the most challenging RNA samples.
How is cDNA synthesis performed in superscript III?
Using the SuperScript® III First-Strand System. cDNA synthesis is performed in the first step using either total RNA or poly(A) +-selected RNA primed with oligo(dT), random primers, or a gene-specific primer. In the second step, PCR is performed in a separate tube using primers specific for the gene of interest.
How does Invitrogen superscript 2 reverse transcriptase work?
Invitrogen SuperScript II Reverse Transcriptase is a genetically engineered MMLV reverse transcriptase (RT) with reduced RNase H activity and increased thermal stability compared to wild-type MMLV RT. Mutations in the RNase H domain of the enzyme eliminate degradation of the RNA during first-strand cDNA synthesis,…
How much RNA can be detected with superscript III?
RNA targets from 100 bp to >12 kb can be detected with this system. The amount of starting material can vary from 1 pg–5 µg of total RNA. SuperScript® III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability.