b"Considerations for selectingthe right 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 10036The ability of a reverse transcriptase to generate cDNA from the lowest34 80t Activity (%)amount of input RNA is an important attribute when working with low-copy32 60Cgenes or difficult sample sources where RNA may have already degraded. 30 10%5%0.0025%40isopropanol BugBuster Formalin28 0.01 U/uL Heparin 20 No inhibitors0.05% Bile salts26 0Thermostability 0 0.5 1.0 1.5 2.0 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 BRlog 10 [degraded RNA] (ng) Time (min)8.0 kbThermostable reverse transcriptases allow reactions to occur at higher6.0 Q SuperScript III enzyme P4.0SuperScript III enzyme3.0temperatures, which help denature RNA with strong secondary structure orSuperScript IV enzyme BL SuperScript IV enzyme BL2.0P 1.5 N Q Nhigh GC content, for generation of longer cDNA, higher cDNA yields, andBR 1.0 T BRbetter coverage of RNA populations 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 without30 M hematin 0.01 U/L heparin 0.5% bile saltsreleasing the template. Highly processive reverse transcriptases allow 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 Qsynthesis of longer cDNA strands in a shorter reaction time, and overall better efficiency in making full-length cDNA. kb SSIV N P T9.023.1 kb6.0 kb 9.48.0Inhibitor tolerance 6.0 6.6Compounds that have inhibitory effects on reverse transcriptases are3.0 4.0 2.33.0 2common in RNA samples even after purification. Their sources include2.01.5reagents used for RNA isolation and contaminants carried over from1.5biological samples. Reverse transcriptases resistant to common inhibitors0.5 1.0help minimize inconsistent or suboptimal results in cDNA-based assays. 0.5(A) Sensitivity, (B) thermostability, (C) processivity, and (D) inhibitor tolerance of reverse transcriptases can affect the quantity and length of cDNA.Find out more at thermofisher.com/reverse-transcription 15"