b"Considerations for selecting theright reverse transcriptaseSensitivity, thermostability, processivity, and inhibitor tolerance of reverse transcriptases all affect the quantity and length of cDNA synthesized. A Degraded Arabidopsis RNAB cDNA synthesis at 50Ctranscription Reverse Target: Gin synthetase40 120Sensitivity 38 100The ability of a reverse transcriptase to generate cDNA from the lowest amount of3680input RNA is an important attribute when working with low-copy genes or difficultt 34 Activity (%) 60Csample sources where RNA may have already degraded. 32 4030 10%5%0.0025% isopropanol BugBuster Formalin28 0.01 U/uL Heparin 20 No inhibitors0.05% Bile saltsThermostability 26 0 0.5 1.0 1.5 2.0 0 0 10 20 30 40 50 60SSIIISSIV N SSIIISSIV BL SSIIISSIV BL BR SSIIISSIV P N T Q SSIIISSIV P N T Q BL SSIIISSIV P N Q T BRThermostable reverse transcriptases allow reactions to occur at higher temperatures,log 10 [degraded RNA] (ng) Time (min)8.0 kb6.04.0 SuperScript III enzyme Pwhich help denature RNA with strong secondary structure or high GC content,SuperScript III enzyme Q SuperScript IV enzyme BL3.0SuperScript IV enzyme BL2.0for generation of longer cDNA, higher cDNA yields, and better coverage of RNAP 1.5 N Q NBR 1.0 T BRpopulations in the cDNA. 0.5C cDNA synthesis in 10 min D cDNA synthesis in the presenceProcessivity of common inhibitorsSuperScript III enzyme SuperScript IV enzyme P T BR Q NDNA/Hind IIIProcessivity is the enzyme's ability to add consecutive nucleotides without releasing 30 M hematin 0.01 U/L heparin 0.5% bile saltsthe template. Highly processive reverse transcriptases allow synthesis of longer SuperScript II enzyme SuperScript IV enzyme P N T Q SuperScript II enzyme SuperScript IV enzyme P N T Q SuperScript II enzyme SuperScript IV enzyme P N T QcDNA strands in a shorter reaction time, and overall better efficiency in making full-length cDNA. kbSSIV N P T9.023.1 kbkbInhibitor tolerance 6.0 8.0 9.46.6Compounds that have inhibitory effects on reverse transcriptases are common6.03.0 4.02.3in RNA samples even after purification. Their sources include reagents used for3.0 22.0RNA isolation and contaminants carried over from biological samples. Reverse1.5 1.5transcriptases resistant to common inhibitors help minimize inconsistent or1.0suboptimal results in cDNA-based assays. 0.5 0.5(A) Sensitivity, (B) thermostability, (C) processivity, and (D) inhibitor tolerance of reverse transcriptases can affect the quantity and length of cDNA.Learn more at thermofisher.com/reverse-transcription Contents 15"