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HIGH-PERFORM A NCE
GENE E XPRESSION
RNA Purification
cDNA Synthesis
Real-Time PCR
I S O L AT E I I
RNA KITS
S ensi FA S T c D N A
SYNTHESIS KIT
S ensi FA S T P R O B E
AND SYBR® KITS
FROM SAMPLE TO RE AL-TIME
RESULTS WITH BIOLINE
BIOLINE PROVIDES HIGH-PERFORMANCE PRODUCTS FOR
ALL OF YOUR GENE EXPRESSION STUDIES.
Start with ISOLATE II Kits for purification and concentration of high-integrity total
RNA from a wide range of starting materials. Continue with the SensiFAST cDNA
Synthesis Kit to generate real-time ready cDNA for complete unbiased 5’ to 3’
RNA sequence representation.
When used in combination with SensiFAST Probe and SYBR® Kits, which use
the latest advances in buffer chemistry, Bioline delivers ultra-sensitive real-time
results in less than an hour.
ISOLATE II RNA KITS
RNA
Purification
TOTAL RNA PURIFICATION
RNA CLEAN-UP AND CONCENTRATION
RNA MINI KIT
Fast, efficient isolation of pure total RNA
from a variety of sources.
R N A M I C RO C L E A N - U P K I T
Purify and concentrate RNA, free from enzymatic
inhibitors or impurities. Innovative column design
allows as little as 5μl elution volumes.
RNA PL ANT KIT
Pure total RNA isolation from plant tissues,
including leaves, bark, roots and fruits.
R N A M I C RO K I T
For isolation of pure total RNA from small
samples, including single cells and biopsy
material.
M1
• Simultaneous RNA clean-up and concentration
• Concentrates RNA up to 50-fold.
• Ideal for RNA purification following enzymatic
digests or phenol-extracts.
2 34 5 67 M
Fig. 1 High-quality RNA isolated from plant tissue
RNA was isolated from 20mg freeze-dried budding leaves of
Arabidopsis thaliana using ISOLATE II RNA Plant Kit. The extracted
RNA was diluted in a 2-fold serial dilution (15ng, lanes 1-7 respectively)
and PCR was performed using MyTaq One-Step RT-PCR Kit.
HyperLadder™ 1kb (M). The results illustrate the quality of the RNA
obtained, as it can be used for very sensitive cDNA synthesis and
PCR without further purification.
Fig. 2 Consistent RNA clean-up
RNA was isolated from HeLa cells using standard phenol
extraction split into eight 22μg samples and purified using
the ISOLATE II RNA Micro Clean-Up Kit. The cleaned
RNA samples were run on an Agilent Bioanalyzer 2100.
All RIN values were >9, indicating highly pure RNA and
ideal for downstream applications.
SensiFAST™ cDNA SYNTHESIS KIT
cDNA
Synthesis
GENERATE REAL-TIME READY
FIRST STRAND cDNA FROM TOTAL
RNA TEMPLATE.
• Reproducible - Unique reverse transcriptase
and new TransAmp buffer generate
consistent first-strand cDNA
• Broad dynamic range - ideal for dilute and
low-copy samples
• Sensitive - real-time RT-PCR analysis from
as little as 1pg of starting total RNA
• Unbiased cDNA synthesis - complete 5’ to
3’ RNA sequence representation
• Rapid - 30 minute cDNA Synthesis protocol,
ideal for high throughput applications
∆Rn
Amplification Plot
Cycle
Fig. 3 SensiFAST cDNA Synthesis kit gives unbiased synthesis
Primer pairs designed at 1kb intervals across the same transcript. Unlike Supplier B
(blue), the SensiFAST cDNA Synthesis Kit (red) has no bias across a 5kb transcript.
Amplification curves
Fluorescence (465-510)
The SensiFAST cDNA Synthesis Kit has been
developed to generate real-time ready first
strand cDNA from total RNA template. The kit
contains all necessary components in the
unique TransAmp™ buffer system to deliver
the highest quality cDNA. An optimized blend
of random hexamer and anchored oligo(dT)
primers provide robust, consistent and
unbiased first-strand synthesis over a broad
range of RNA template concentrations, making
SensiFAST cDNA Synthesis Kit the ideal
partner for SensiFAST Probe and SYBR® realtime PCR kits.
100ng
100pg
Cycles
Fig. 4 Reproducibility of SensiFAST cDNA Synthesis Kit
SensiFAST cDNA Synthesis Kit was employed in 48 independent first-strand reactions
in triplicate, containing 100ng or 100pg of total RNA. The results demonstrate the
excellent reproducibility of SensiFAST cDNA Synthesis Kit (the same Ct values), across
the 144 wells with high levels and the 144 wells with low levels of input target mRNA.
SensiFAST™ PROBE AND SYBR ® KITS
Real-Time
PCR
REAL-TIME RESULTS IN LESS
THAN AN HOUR.
SensiFAST uses the latest developments in
real-time PCR to realize the fastest cycling
times and greatest sensitivity, without
compromising accuracy, reproducibility or
performance.
• Fast - optimized for fast cycling conditions
(under 30 minutes)
• High specificity - leading to better efficiency
and more confidence in your results
• Ultra-sensitive - perfect for low-copy
number samples
• Universal - compatible with all standard and
fast cycling instruments.
A.
B.
Fig. 7 Superior performance in multiplex assays
A 10-fold serial dilution of human cDNA was amplified with 4 different probes
(results displayed for a single gene), both in singleplex reactions (blue line) and
quadruplex reactions (red line). The results illustrate that SensiFAST Probe No-ROX
has high sensitivity and excellent reproducibility for both singleplex and multiplex
reactions (Fig. 7A) and no reduction of efficiency (Fig. 7B) that is often associated
with multiplexing. There is no change in Ct values between singleplex and multiplex
reactions.
Fig. 5 Superior speed and specificity under fast cycling conditions
A 10-fold serial dilution of human genomic DNA was amplified using SensiFAST
Probe Lo-ROX (red line) and the results were compared with amplifications
using a kit from Supplier B (blue line) under fast cycling conditions (45 cycles of
95°C for 10s, 60°C for 10s). The results illustrate the quality of SensiFAST Probe
Lo-ROX, which is faster (earlier Ct) and more sensitive than Supplier B.
Fig. 6 High sensitivity, enabling detection of expressed genes
10-fold serial dilutions of human genomic DNA (in triplicate) over 5 orders of
magnitude was amplified using SensiFAST Probe No-ROX (red line) and the results
were compared with amplifications using a kit from Supplier R (blue line). Results
illustrate that SensiFAST Probe is at least 10 fold more sensitive in this assay.
“ ”
HERE IS WHAT OTHER
RESE ARCHERS SAY
ABOUT OUR PRODUCTS
“SensiFAST ™ consistently exhibits excellent sensitivity
in our differential gene expression assays and superior
efficiency compared with our previous qPCR mix.”
National Institutes of Health
Bethesda, Maryland, USA
FOR MORE INFORMATION PLEASE VISIT
bioline.com/genex
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