Document Number : F701-28-52-E

Rev. No. : 1.0

Rev. Date : 2024.02.26
Issue Date : 2023.08.17

ADPSTM Smart DNA Polymerase II

For Research Use Only

Contents Quality control

Cat No. Smart DNA 5X ADPS Buffer 4 dNTP Mix ▪ Specific activity: ≥100,000 U/mg
Pol. II (5U/μL) (Each 10 mM) ▪ Allele discrimination assay: Meet to acceptance criteria
8P01BAA 250 Units, 0.05 ml 2 x 1 ml 0.25 ml ▪ Physical purity: ≥95% in SDS-PAGE
▪ DNase, RNase activity: None detected
8P01BBA 500 Units, 0.1 ml 4 x 1 ml 0.5 ml
▪ Endonuclease, Exonuclease activity: None detected
8P01BCA 1,000 Units, 0.2 ml 8 x 1 ml 1 ml ▪ E. coli DNA contamination: None detected
▪ Bioburden assay: None detected

Standard PCR reaction

Description

Allele specific Primer Probe

ADPS™ Smart DNA Polymerase II is an improved version of DNA Template
polymerase derived from Thermus aquaticus. This enzyme exhibits distinct F Q
5’:::::::::C :::::::::
features, distinguishing between matching and mismatching at the 3' end of 3’ G 5’
::::::::
5’
primers. In cases of mismatch, it demonstrates significantly reduced
Variation point Common Primer
amplification efficiency compared to wild-type Taq polymerase. The unique
characteristics of ADPS™ Smart DNA Polymerase II, along with an
optimized buffer system, enable high-performance allele-specific PCR PCR mixture (20-μL reaction)
without the need for special modifications during the PCR process. Except
for allele discrimination ability, all characteristics of the enzyme are the same Components Volume Final conc.
as those of general Taq polymerase, including polymerase activity and 5’ to 5X ADPS buffer 4 4 μL
3’ exonuclease activity, but it lacks 3’ to 5’ exonuclease activity. The utility dNTP Mix (Each 10 mM) 0.5 μL Each 250 μM
of ADPS™ Smart DNA Polymerase II extends across a wide range of 10 μM Allele-specific primer 0.5-1 μL 250 - 500 nM
applications, including SNP genotyping, genetic disease testing, and other
10 μM Common primer 0.5-1 μL 250 - 500 nM
genetic alterations. Its versatility spans from variant typing requiring high-
10 μM Probe (for Real-Time PCR only) 0.5-1 μL 250 - 500 nM
sensitivity to ultra-sensitive detection of rare mutations, such as those
Template Variable 0.1 - 500 ng
associated with cancer genes.
Smart DNA polymerase II (5 U/μL) 0.1-0.2 μL 0.5-1 U
Nuclease-Free Water Up to 20 μL
ADPS (Allele-Discriminating Priming System)
PCR cycle
Match
Step Temp. Time Cycle
5’ C 5’ C
Initial Denaturation 94℃ 5 min 1
3’ G 3’ G
Denaturation 94℃ 10-20 sec
Annealing 60℃* 10-30 sec 25-40
WT Taq Smart Pol
Extension 72℃ 10-30 sec
No Final Extension 72℃ 5 min 1
5’ C 5’ C extension
X X *Annealing temperature should be optimized for each primer set based on the primer Tm.
3’ T 3’ T
Mismatch * Compatible Real-Time PCR System

• Applied Biosystems 7500, 7500 Fast, QuantStudio3 Real-Time PCR

Storage and Stability
System
• Bio-Rad CFX96TM, CFX384TM Real-Time PCR Detection System
When stored at -15 to -25°C, the product is stable until the
• Roche LightCycler® 480 System
expiration date printed on the label.
• Qiagen Rotor-Gene Q
Avoid repeated freeze/thaw cycles (> 5 times).
• Other Real-Time PCR equipment using fluorescent signal

Applications
• Allele-Specific PCR (Conventional or Real-Time PCR)
• SNP genotyping
• Multiplex PCR
• Ultrasensitive detection of rare mutations
• Methylation Specific PCR
General considerations for allele-specific PCR

The optimal conditions (component concentrations and cycle conditions)

vary according to target variation types, amplicon size, reaction volumes,
and type of thermal cycler used and must be determined for each individual
experimental system.
• The allele-specific nucleotide should ideally be the last nucleotide at
the 3′ end of the primer. To increase allele discrimination sensitivity,
additional mismatches near the 3' end can be introduced.
• Allele specific primers generally range in length from 15-23 bases and
have a melting temperature about 60℃.
• Common primers have a melting temperature about 63℃.
• Primer sequences should exhibit a 40-60% GC content, and one should
avoid sequences that could generate internal secondary structures.
• The optimal enzyme concentration range 0.5-1 U/20 μL and can be
increased in cases of low PCR yield.
• The optimal concentration of each dNTP should be between 50 and 500
µM, and it is recommended to use at concentration of 250 µM dNTP.
• The primer and probe concentrations should be optimized within the
range of 100 nM to 500 nM. We recommend using 200 nM for the
primer and 400 nM for the probe in a 20 μL Real-Time PCR reaction.

Reference

Lim, Y., Park, I. H., Lee, H. H., Baek, K., Lee, B. C., & Cho, G. (2022).
Modified Taq DNA Polymerase for Allele-Specific Ultra-Sensitive
Detection of Genetic Variants. The Journal of Molecular Diagnostics,
24(11), 1128-1142.

Park, I. H., Son, D. S., Choi, Y. L., Choi, J. H., Park, J. E., Jeon, Y. J., ... &
Kim, J. (2023). Clinical Validation of the Unparalleled Sensitivity of the
Novel ADPS Technology-Based EGFR Mutation Assay in Patients with
Operable Non-Small Cell Lung Cancer. Cancer Research and Treatment.
56(1), 81-91.

Precautions and Disclaimer

This product is for Research use only. Please consult the Material Safety
Data Sheet for information regarding hazards and safe handling practices.

Contact and Support

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