UDGin™ PCR Cleaner Mix, 20X

Applications


  • Efficient prevention of carry-over contamination in qPCR

  • Efficient prevention of carry-over contamination in PCR

Benefits


  • Time saving ready-to-use mix with UDG and dUTP

  • Compatible with all highQu qPCR and PCR master mixes

Catalog. # Product Name Size Price Qty
UDG0101 UDGin™ PCR Cleaner Mix, 20X 500 r of 20 µl
€49.00

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Description

UDGin™ PCR Cleaner Mix is an efficient tool for carry-over contamination prevention in qPCR or PCR. It is optimized for the use in conjunction with highQu qPCR or PCR products.

UDGin™ PCR Cleaner Mix, is a ready to use 20X mix of optimally combined Uracil-DNA Glycosylase (UDG) and dUTP in a buffer with stabilizers. 1 µl of UDGin™ PCR Cleaner Mix added into each 20 µl PCR reaction prevents amplification of DNA carried over from previous PCR performed with dUTP.

During 10 minutes incubation before the PCR start, UDG hydrolyzes the N-glycosilic bond between uracil and sugar leaving apyrimidinic sites in uracil containing DNA which is then cleaved by heat during the first PCR cycle, thus only the newly added template is amplified. UDG is inactivated during the initial PCR denaturation step and does not destroy newly synthesized dUTP containing DNAs. PCR products obtained when UDGin™ PCR Cleaner Mix was used contain uracil, therefore they will be destroyed again by UDG before the next PCR start.

 

Use of UDG in certain countries for certain applications may be covered by patents and may require a license.

 

 

Protocols

 

Print version of the protocol: Product Insert UDGin™ PCR Cleaner Mix, 20X

Protocol:
  • Follow the protocol of ORA™ qPCR Mix or ALLin™ PCR Enzyme or Mastermix that you use. Only three additional steps are generally required for preventing carry-over contamination in PCR using UDGin™ PCR Cleaner Mix:

1. Before adding water into PCR reactions, add 1µl of UDGin™ PCR Cleaner Mix, 20X into each 20 µl reaction. Use less water accordingly.
2. Set your qPCR/PCR instrument to perform at the beginning one cycle of 37°C incubation for 10 min.
3. After incubation always perform one cycle of longer initial denaturation (and UDG inactivation) for 5 minutes at 95°C.

 

Note:

  • Store PCR reactions on ice.
  • Before subsequent applications, consider that your PCR products contain uracil. This has no influence on electrophoresis and sequencing, but might affect the cleavage with certain restriction enzymes (check enzyme performance on U containing sites). When cloning, use only ung- bacterial hosts for transformations.

 

Prepare a 20 µl reaction:

 

Reverse Primer 100 - 400 nM final c.
Forward Primer 100 - 400 nM final c.
cDNA Template or
gDNA Template
<100ng  or
<1 μg
UDGin™ PCR Cleaner Mix, 20X 1 μl (to final 1X conc.)
PCR Water to 10 μl
ORA™ qPCR Green ROX L Mix, 2X 10 μl

 

  • Mix gently, avoid bubbles.
  • Place into the instrument (SYBR® Green or FAM channel), set like:

 

UDG treatment 1 cycle: 37°C - 10 min
(uracil-containing DNA hydrolysis)
Initial denaturation 1 cycle: 95°C - 2 min for cDNA, or
1 cycle: 95°C - 3 min for gDNA
Denaturation 40 cycles: 95°C - 5 sec
Annealing/extension 40 cycles: 60-65°C - 20-30 sec

 

  • Follow instrument instructions for melt curve analysis.

Specifications

Cat.

Size

Components

Composition

UDG0101 500 r of
20 µl
0.5 ml - UDGin™ PCR Cleaner Mix, 20X 20X master mix for PCR carry-over contamination prevention, contains optimized concentration of dUTP, Uracil DNA Glycosylase (UDG), and stabilizers.

Storage

  In the dark at -20°C.

Resources

 

Product Insert