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Molecular Cloning Laboratories
MCNextTM
SYBR®
Fast qPCR Library Quantification Kit
- The Revolution is Here
Molecular Cloning Laboratories
Molecular Cloning Laboratories
The Only Ngs Library Quant Kit Using
A Library Standard
Simpler workflow – direct (MC kit) vs. indirect (KAPA kit)
cluster density conversion
Broader dynamic range of measurement – 7-log (MC kit)
vs. 6-log (KAPA kit)
Superior accuracy – Phi X library (MC kit) vs. single
DNA fragment (KAPA kit) as standard
Reliable – more consistent in quantification comparing
with KAPA kit
Molecular Cloning Laboratories
Kit Components
Component name Volume
2x MCNext SYBR Fast qPCR Master Mix 5 mL
10x Primer Mix 1 mL
Phi X Library Standards 10 µL
Storage Buffer 3 mL
50x Low ROX (optional) 200 µL
50x High ROX (optional) 200 µL
Molecular Cloning Laboratories
Kit Types
Kit type Description Cat #
Universal kit
No ROX added to 2x
qPCR Master Mix
IQPQ-UN
Low ROX kit
Low concentration
ROX in 2x qPCR
Master Mix
IQPQ-LR
Regular ROX kit
Regular concentration
ROX in 2x qPCR
Master Mix
IQPQ
Molecular Cloning Laboratories
Broad Compatibility Coverage
Reference Dye Compatible Instruments
No ROX
BioRad iCycler MiniOpticon, Opticon 2, iQ5; Roche LightCycler
480; MJ Research DNA Engine Opticon 2, Chromo 4; Corbett
Rotogene 3000, 6000
Low ROX
ABI 7500 qPCR Systems, ViiA 7, QuantStudio 12K Flex, Agilent
Mx3000P and Mx4000
Regular ROX
ABI PRISM 7000, 7700, 7900HT, ABI 7300 qPCR Systems, Ge-
neAmp 5700, StepOne, and the StepOnePlus
Molecular Cloning Laboratories
Higher Signal Strength and Better Efficiency
for Library Quantification
Higher fluorescence signal
Better efficiency
1.000
1.000 E+1
Cycle
10 15 20 25 30 35
MCNext Library qPCR Quantification Kit
KAPA SYBR FAST qPCR Kit®
Molecular Cloning Laboratories
Broader Dynamic Range for Library
Quantification
10x better sample
quantification range
Better efficiency
Broader dynamic range
MCNext Library qPCR Quantification Kit
KAPA SYBR FAST qPCR Kit
0
5
10
15
20
25
30
-4 -3 -2 -1 0 1 2
Ct
y = -3.426x + 13.67
R² = 0.9997
y = -3.423x + 14.078
R² = 0.9998
®
Molecular Cloning Laboratories
Superior Accuracy for Library Quantification
Electropherogram
from Agilent
2100 Bioanalyzer
indicates library
average size of
500bp size
The only kit on the world wide market using an NGS library to quantify NGS
libraries
KAPA kit uses a 525 bp single cDNA fragment to quantify NGS libraries
Molecular Cloning Laboratories
High Quality of the Library Quantification
Standards
Melting curve of library standards after
PCR amplification
Single peak
No nonspecific amplification products
6.900 E-2
5.900 E-2
4.900 E-2
3.900 E-2
2.900 E-2
1.900 E-2
9.000 E-3
60.0 65.0 70.0 75.0 80.0 85.0 90.0
Derivative
Temperature (˚C)
Molecular Cloning Laboratories
Better Quantification Consistency than
KAPA kit
* Actual Illumina Phi X library concentration = 10 nM

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MCNext Sybr qPCR quantification Kit

  • 1. Molecular Cloning Laboratories MCNextTM SYBR® Fast qPCR Library Quantification Kit - The Revolution is Here Molecular Cloning Laboratories
  • 2. Molecular Cloning Laboratories The Only Ngs Library Quant Kit Using A Library Standard Simpler workflow – direct (MC kit) vs. indirect (KAPA kit) cluster density conversion Broader dynamic range of measurement – 7-log (MC kit) vs. 6-log (KAPA kit) Superior accuracy – Phi X library (MC kit) vs. single DNA fragment (KAPA kit) as standard Reliable – more consistent in quantification comparing with KAPA kit
  • 3. Molecular Cloning Laboratories Kit Components Component name Volume 2x MCNext SYBR Fast qPCR Master Mix 5 mL 10x Primer Mix 1 mL Phi X Library Standards 10 µL Storage Buffer 3 mL 50x Low ROX (optional) 200 µL 50x High ROX (optional) 200 µL
  • 4. Molecular Cloning Laboratories Kit Types Kit type Description Cat # Universal kit No ROX added to 2x qPCR Master Mix IQPQ-UN Low ROX kit Low concentration ROX in 2x qPCR Master Mix IQPQ-LR Regular ROX kit Regular concentration ROX in 2x qPCR Master Mix IQPQ
  • 5. Molecular Cloning Laboratories Broad Compatibility Coverage Reference Dye Compatible Instruments No ROX BioRad iCycler MiniOpticon, Opticon 2, iQ5; Roche LightCycler 480; MJ Research DNA Engine Opticon 2, Chromo 4; Corbett Rotogene 3000, 6000 Low ROX ABI 7500 qPCR Systems, ViiA 7, QuantStudio 12K Flex, Agilent Mx3000P and Mx4000 Regular ROX ABI PRISM 7000, 7700, 7900HT, ABI 7300 qPCR Systems, Ge- neAmp 5700, StepOne, and the StepOnePlus
  • 6. Molecular Cloning Laboratories Higher Signal Strength and Better Efficiency for Library Quantification Higher fluorescence signal Better efficiency 1.000 1.000 E+1 Cycle 10 15 20 25 30 35 MCNext Library qPCR Quantification Kit KAPA SYBR FAST qPCR Kit®
  • 7. Molecular Cloning Laboratories Broader Dynamic Range for Library Quantification 10x better sample quantification range Better efficiency Broader dynamic range MCNext Library qPCR Quantification Kit KAPA SYBR FAST qPCR Kit 0 5 10 15 20 25 30 -4 -3 -2 -1 0 1 2 Ct y = -3.426x + 13.67 R² = 0.9997 y = -3.423x + 14.078 R² = 0.9998 ®
  • 8. Molecular Cloning Laboratories Superior Accuracy for Library Quantification Electropherogram from Agilent 2100 Bioanalyzer indicates library average size of 500bp size The only kit on the world wide market using an NGS library to quantify NGS libraries KAPA kit uses a 525 bp single cDNA fragment to quantify NGS libraries
  • 9. Molecular Cloning Laboratories High Quality of the Library Quantification Standards Melting curve of library standards after PCR amplification Single peak No nonspecific amplification products 6.900 E-2 5.900 E-2 4.900 E-2 3.900 E-2 2.900 E-2 1.900 E-2 9.000 E-3 60.0 65.0 70.0 75.0 80.0 85.0 90.0 Derivative Temperature (˚C)
  • 10. Molecular Cloning Laboratories Better Quantification Consistency than KAPA kit * Actual Illumina Phi X library concentration = 10 nM