Genoprice :
PCR solution
EzWayTM Contam-Free PCR Solution
1. Catalog No. K33100
2. Quantity 1ml
3. Introduction Removing of DNA contaminants is the most important factor for
the accurate PCR
(Polymerase Chain Reaction) result. Even the trace amount of DNA contaminants
can
serve as templates, thus it results in amplification of the wrong template and
leads to a
false positive result. Ensuring that the previous PCR products (carry-over
contamination)
are not reamplified in subsequent PCR amplification is very important especially
to a
laboratory that performs PCR as a routine work such as clinical diagnosis or
environmental monitoring.
EzWayTM Contam-Free PCR Solution is designed to eliminate DNA contaminants
before
PCR performance using a photochemical which is known as an useful agent to
destroy
DNA's ability that acts like a template for PCR. This photochemical is
intercalated into
double-stranded nucleic acids and form a covalent interstrand crosslink after
photoactivation with UV light. Therefore, it is useful to extinguish the
template activity of
contaminating DNAs.
Easy to use: Just mix with your PCR premix and then exposure to UV light
Prevention of contamination: Elimination of the DNA contaminants
High efficiency of the PCR: No-false positive
4. Storage & Stability Store at room temperature. (Keep in the dark.)
5. Protocol 1. Prepare PCR master mixture of dNTPs, oligonucleotide primers and
reaction buffers,
except Taq enzyme and template DNA.
2. Add 1 ul of EzWayTM Contam-Free PCR Solution to 100 ul of PCR master mixture.
Note: This solution is supplied as 100x concentration. Use it as it is without
any dilution.
3. Incubate at 4°C for 30 minutes.
4. Exposure the mixture to UV light with concentration of 1 J / 200 ul with the
UV
Linker. For example, if you make 400 ul of PCR master mixture, irradiate 2 J of
UV
to sample.)
5. Cool the UV treated tube on ice.
6. Add Taq polymerase and mix well.
7. Aliquot PCR master mix which was prepared at the Step 6 to the individual PCR
tubes and apply template DNA to each tube.
8. Perform PCR amplification.
Lane N: No template
Lane S: Plus Template
Lane M: DNA Ladder
Lane 1-8 (Panel A): Treatment of
EzWayTM Contam-Free PCR
Solution plus UV Irradiation
Lane 9-10 (Panel B): No
treatment of either EzWayTM
Contam-Free PCR Solution nor
UV Irradiation
Efficiency
False-positive results for the no-template controls (Lane N in Panel B) under no
treatment of
EzWayTM Contam-Free PCR Solution were shown. No false-positive signals were
detected when
the master mixture was treated with EzWayTM Contam-Free PCR Solution and UV
irradiation as
shown in Panel A. According to this experiment, UV exposure times were optimized
in the range of
10 min. Excessive UV exposure time might reduce PCR sensitivity.
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