THUNDERBIRD Next SYBR® qPCR Mix

Product#: QPX-201
$1.00
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THUNDERBIRDTM Next SYBR® qPCR Mix


Cat. No. List below

 

Product Name

Cat. No.

Size

Storage

Price USD ($)

Trial

QPX-201_Trial

1 mL 

-20 oC

1.00

THUNDERBIRDTM Next SYBR® qPCR Mix

QPX-201_1PK

1.67 mL x 3 tubes

173.00

THUNDERBIRDTM Next SYBR® qPCR Mix QPX-201_5PK (1.67 mL x 3 tubes) x5 791.00

Thunderbird_Comparison
Description


THUNDERBIRDTM Next SYBR® qPCR Mix is a highly efficient 2× Master Mix for real-time PCR using SYBR® Green I. The master mix contains all required components, except primers. The master mix facilitates reaction setup, and improves the reproducibility of experiments.

This product is an improved version of THUNDERBIRDTM SYBR® qPCR Mix (Code No. QPS-201). In particular, the reaction specificity and PCR efficiency is enhanced.

 

  • High amplification efficiency
  • High sensitivity of Low copy areas
  • High specificity
  • Corresponding to high-speed cycle (10 sec. extension)
  • Improving the stability of mixed reaction solution
  • Prevention of false positives due to carry-over, (combined with separately available UNG)
  • Broad instrument compatibility
  • Colored master mix

Features
 
Feature 1: High amplification efficiency
THUNDERBIRD Next SYBR qPCR Mix allows efficient amplification up to 500bp with minimal variability in PCR efficiency per target.
Using artificial gene as template and forward primer as common, reverse primer
was designed so that the amplification product lengths were 100bp, 200bp,
300bp, 400bp and 500bp. We used these primers to amplify 107 to 10
copies using THUNDERBIRD Next SYBR qPCR Mix and other companies to
compare the PCR efficiency for each amplification length. As a result, the
amplification efficiency may decrease or be undetectable as the target length
increases in other companies, but stable PCR efficiency was achieved with
THUNDERBIRD Next SYBR qPCR Mix.
QPX-201_F-1
 
Feature 2: High sensitivity of Low copy areas
High efficiency and specific amplification to low copies is possible, allowing analysis with a wide measurement range.
If one copy target can be amplified, the number of detected copies would be
equivalent to the expected number of detected copies from the Poisson distribution.
When one copy is added, the theoretical value from the Poisson distribution is
37% of the probability of having 0 copies and 63% of the probabilityof having
one or more copies. 
THUNDERBIRD Next SYBR qPCR Mix was used to detect 96 samples using Salmonella genome diluted to one copy as a template.

The results showed that 38.5% of the samples were undetected and 61.5% were detected, which is equivalent to the expected number of samples from the Poisson
distribution, suggesting that one copy equivalent can be detected.
QPX-201_F-2
 
Feature 3: High specificity
Reducing non-specific reactions improves the reliability of detecting lowconcentration targets.

cDNA from total RNA of Hela cells synthesized by reverse-transcription reagents (Code No. FSQ-101) was used to amplify a 65-bp amplified G3PDH at 5-fold dilutions of cDNA. As a result, nonspecific amplification occurred in the low copy range in other companies, but nonspecific amplification was not observed by using THUNDERBIRD Next SYBR qPCR Mix, allowing accurate quantification to the low copy range.
 
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Feature 4: Corresponding to high-speed cycle
THUNDERBIRD Next SYBR qPCR Mix can also be applicable by high-speed PCR with an extension time of 10 seconds.

Using cDNA from Hela cells total RNA synthesized by reverse-transcription reagents (Code No. FSQ-101), amplification of b-actin gene (316bp) was amplified by "normal cycle with 30sec. extension " and "high-speed cycle with 10sec. extension".
As a result, other company products that recommend normal cycling could not be amplified efficiently by high-speed cycling, but  THUNDERBIRD Next SYBR qPCR Mix could be amplified efficiently by high-speed cycling.

 
 
Feature 5: The stability of mixed solution
Stable results are obtained due to high stability when primer and templates are mixed.

Primers and template (cDNA from Hela cellular total RNA) was mixed into the PCR-reaction solution, and amplification of the
target was performed immediately or after standing for 48 hours at a light-shielding room temperature.
As a result, Ct values decreased after 48 hours in conventional THUNDERBIRD and other products. But in THUNDERBIRD
Next SYBR qPCR Mix, Ct values remained stable even after 48 hours.


Differences with ΔCt greater than or equal to 0.5 are shown in yellow.
 
Feature 6: Prevention of false positives
The products include dUTP.
Therefore, false positives due to carry-over contamination can be prevented by using Uracil-DNA Glycosylase (UNG).

To confirm UNG treatment for preventing carry-over contamination.
PCR products containing dUTP (104 copies) were used as template, THUNDERBIRD Next SYBR qPCR Mix and Uracil- DNA Glycosylase(UNG), Heat-labile (Code No. UNG-101) were added, and amplification of the same was performed by real-time PCR.
As a result, we were able to confirm that the first PCR products were degraded by UNG treatment completely.

 
 
Feature 7: Broad instrument compatibility

QPX-201 already includes universal passive reference dye.
As QPX-201 is already mixed with universal reference dye, any instrument can be used with same reagent. Therefore, it is not necessary to mix ROX and use some reagents according to the real-time PCR instrument.
 
QPS-201 need to mix ROX passive reference dye separately.
  • 1×ROX required: ABI 7900, ABI StepOne, ABI StepOnePlus, ABI ViiA7
  • 0.1×ROX required: ABI 7500Fast, ABI QuantStudio
  • ROX is not required: Roche, Bio-Rad, Agilent, TaKaRa, Qiagen
QPK-201 includes 1×passive reference dye.
QPK-201 cannot use for 0.1×ROX required qPCR instruments such as ABI 7500Fast, QuantStudio.

Example of available devices
 Applied
 Biosystems
 7300 / 7500 / 7500 Fast / StepOne /
 StepOne PlusViiA 7 / QuantStudio
 Roche
 Diagnostics
 LightCycler 1. x / 2.0 / Nano / 96 / 480
 Bio-Rad/MJ  MiniOpticon / CFX96 Touch
 Agilent
 Technologies
 Mx3000 / Mx3005P / Mx4000 / AriaMx
 TaKaRa  Dice / DiceII / Dice Lite / Dice III
 QIAGEN  Rotor-Gene Q
 BioFlux  Line Gene
 
Feature 8: Colored master mix
The composition of this product contains blue dye.
The dispensed wells of this product are blue, so dispensing errors can be reduced.
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