The OneTaq One-Step RT-PCR Kit offers sensitive and robust end-point detectionof RNA templates. cDNA synthesis and PCR amplification steps are performed ina single reaction using gene-specific primers, resulting in a streamlined RT-PCRprotocol.The kit combines three optimized mixes: OneTaq One-Step Enzyme Mix,OneTaq One-Step Reaction Mix and OneTaq One-Step Quick-Load ReactionMix. The Enzyme Mix combines ProtoScript II Reverse Transcriptase, MurineRNase Inhibitor and OneTaq Hot Start DNA Polymerase. ProtoScript II ReverseTranscriptase is a mutant M-MuLV reverse transcriptase with reduced RNaseH activity and increased thermostability. OneTaq Hot Start DNA Polymerase ismixture of a Hot Start Taq DNA Polymerase combined with a proof-reading DNApolymerase, resulting in high-yield amplification with minimal optimization. TheOneTaq One-Step RT-PCR Kit is capable of amplifying long transcripts up to 9 kbin length.Two optimized reaction mixes are included, OneTaq One-Step Reaction Mix andQuick-Load OneTaq One-Step Reaction Mix. The reaction mixes offer robustconditions for both cDNA synthesis and PCR amplification. The unique OneTaqQuick-Load One-Step Reaction Mix contains additional dyes, offering colorindication for reaction setup as well as direct gel loading.Both total RNA and mRNA can be used as template. The kit can detect a GAPDHtarget as low as 0.1 pg per reaction. It can routinely detect RNA targets up to9 kb. The OneTaq One-Step RT-PCR Kit is capable of multiplex detection of twoor three targets.Figure 1. Detection of RNA templates of different length.About 100 ng of Jurkat total RNA was used in 50 μl reactions following the standard protocol. The target sizeswere Lane 1: 0.7 kb, Lane 2: 1.1 kb, Lane 3: 1.9 kb, Lane 4: 2.3 kb, Lane 5: 2.5 kb, Lane 6: 5.5 kb, Lane 7: 7.6 kband Lane 8: 9.3 kb. The marker lane (M) contains Quick-Load 1 kb Plus DNA Ladder (NEB #N0469).Figure 2. Sensitive detection of RNA down to 1 pg.Serial dilution of HeLa total RNA (Lane 1: 1 μg, Lane 2: 100 ng, Lane 3: 10 ng, Lane 4:1 ng, Lane 5: 100 pg, Lane 6: 10 pg, Lane 7: 1 pg, Lane 8: 0.1 pg) was used in 50 μlreactions following the Standard Protocol. The marker lane (M) contains Quick-Load 1 kb Plus DNA Ladder (NEB #N0469).
This product is related to the following categories:
RT-PCR Products,
Reverse Transcriptases & RT-PCR Products,
cDNA Synthesis & Reverse Transcriptases Products
This product can be used in the following applications:
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