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KAPA SYBR Fast One-Step qRT-PCR Kits

For Research Use Only. Not for use in diagnostic procedures.

KAPA qPCR

Overview

KAPA SYBR FAST One-Step qRT-PCR Kits are designed for optimal performance in stringent qRT-PCR reaction conditions, exhibiting improvements in sensitivity, specificity, reaction efficiency, and fluorescence. The kits contain an M-MuLV reverse transcriptase, an RNase inhibitor, and a novel DNA polymerase engineered through our directed evolution technology.

Benefits of KAPA SYBR FAST One-Step qRT-PCR Kits

  • Rapid one-step, one-tube RNA quantification, reducing experimental variation and contamination
  • A convenient qRT-PCR protocol

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Product highlights

Increase signal and reaction efficiency

  • Consistently higher amplification efficiencies *
  • Higher fluorescence, earlier Cq  values and improved reaction efficiencies compared to other options*

View data showing increased signal and reaction efficiency using KAPA SYBR FAST One-Step qRT-PCR Kit compared to a competitor kit.

 

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Figure 1. RRMI gene (94 bp, 45.7% GC) amplified from 10-fold dilution series of RNA (100 ng to 10 pg per 20 µL reaction) isolated from human placenta. KAPA SYBR FAST One-Step qRT-PCR Kit demonstrates earlier Cq values, higher fluorescence, and optimal reaction efficiencies.

Figure 1 Improved sequencing performance

Figure 2. RRMI gene (94 bp, 45.7% GC) amplified from 10-fold dilution series of RNA (100 ng to 10 pg per 20 µL reaction) isolated from human placenta. KAPA SYBR FAST One-Step qRT-PCR Kit demonstrates earlier Cq values, higher fluorescence, and optimal reaction efficiencies.

Figure 2 RRMI gene (94 bp, 45.7% GC) amplified

Figure 3. RRMI gene (94 bp, 45.7% GC) amplified from 10-fold dilution series of RNA (100 ng to 10 pg per 20 µL reaction) isolated from human placenta. KAPA SYBR FAST One-Step qRT-PCR Kit demonstrates earlier Cq values, higher fluorescence, and optimal reaction efficiencies.

Figure 3 high positive predictive values (PPVs)
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Amplify across a broad range of target lengths

  • Robust performance independent of amplicon size

View data showing high qPCR performance using KAPA SYBR FAST One-Step qRT-PCR Kit across varying lengths of RNA targets.

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Figure 4. MS2 bacteriophage RNA targets of increasing length were amplified at 0.8 pg per 20 µL reaction. KAPA SYBR FAST One-Step qRT-PCR Kit maintained high performance across all amplicon lengths.

Figure 4 MS2 bacteriophage RNA targets of increasing length

Figure 5. MS2 bacteriophage RNA targets of increasing length were amplified at 0.8 pg per 20 µL reaction. KAPA SYBR FAST One-Step qRT-PCR Kit maintained high performance across all amplicon lengths.

Figure 5. MS2 bacteriophage RNA targets

Figure 6. MS2 bacteriophage RNA targets of increasing length were amplified at 0.8 pg per 20 µL reaction. KAPA SYBR FAST One-Step qRT-PCR Kit maintained high performance across all amplicon lengths.

Figure 6. MS2 bacteriophage RNA targets of increasing length
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Improve sensitivity and reproducibility

  • Accurate interrogation across a wide range of RNA template concentrations

View performance data for KAPA SYBR FAST One-Step qRT-PCR Kit showing high and uniform efficiency across varying dilutions of target.

Figure 7. 279 bp fragment amplified in replicates of 6 from a 10-fold dilution series of commercial MS2 bacteriophage RNA (80 pg to 0.8 fg per 20 µL reaction). Reaction efficiency of 95% calculated for all 6 replicates per data.

Figure 7. 279 bp fragment amplified

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*Data on file.

Specifications

  • Starting Material

    RNA

  • Input Amount

    1 pg–1 µg total RNA, 10 fg–100 ng poly(A)RNA, 10-1x108 copies viral RNA

Kits can be stored for up to 6 months at -20˚C.

Kits include KAPA RT Mix, dUTP, and KAPA SYBR FAST qPCR Master Mix, which contains KAPA SYBR FAST DNA Polymerase, reaction buffer, dNTPs, SYBR Green I dye, and MgCl2 at a final concentration of 2.5 mM. Where noted, Master Mixes contain instrument-specific reference dyes, while the Universal kit includes 50X ROX High and 50X ROX Low separately.

Please reference the Instrument Compatibility Chart for guidance on compatible platforms.

Components

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   Kit code  50X KAPA SYBR FAST ROX High 50X KAPA SYBR FAST ROX Low 2X KAPA SYBR FAST qPCR Master Mix (Universal) 2X KAPA SYBR FAST qPCR Master Mix (with equipment-specific dye) 50X KAPA RT Enzyme Mix 10mM dUTP
Universal KK4650 X X X   X X
KK4651            
KK4652            
Bio-Rad iCycler KK4670       X X X
KK4671            
KK4672            
Optimized for Roche LightCycler® 480 KK4680       X X X
KK4681            
KK4682            

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Kits include KAPA RT Mix, dUTP, and KAPA SYBR FAST qPCR Master Mix, which contains KAPA SYBR FAST DNA Polymerase, reaction buffer, dNTPs, SYBR Green I dye, and MgCl2 at a final concentration of 2.5 mM. Where noted, Master Mixes contain instrument-specific reference dyes, while the Universal kit includes 50X ROX High and 50X ROX Low separately.

Component

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KAPA Code
 Roche Cat. No
 Description
 Kit Size
 How to buy
KK4650
 07959613001
 100 x 20 µL reactions (Universal) 1 mL
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KK4651
 07959621001
 500 x 20 µL reactions (Universal) 5 mL
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KK4652
 07959630001
 1000 x 20 µL reactions (Universal) 10 mL
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KK4670
 07959702001
 100 x 20 µL reactions, with fluorescein (Bio-Rad® iCycler™) 1 mL
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KK4671
 07959729001
 500 x 20 µL reactions, with fluorescein (Bio-Rad iCycler) 5 mL
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KK4672
 07959737001
 1000 x 20 µL reactions, with fluorescein (Bio-Rad iCycler) 10 mL
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KK4681
 07959761001
 500 x 20 µL reactions (optimized for use with Roche LightCycler 480) 5 mL
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