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Instruction manual Uracil-DNA Glycosylase (UNG), Heat-labile 1907

F1715K

Uracil-DNA Glycosylase (UNG),Heat-labile
Code No. UNG-101 200 U×1
Store at －20°C[ 1 ] Introduction
Uracil-DNA Glycosylase (UNG) hydrolyzes the N-glycosylic bond between the deoxyribose
sugar and the base in uracil-containing DNA leaving an apyrimidinic site in DNA. Since DNA with
an apyrimidinic site is degraded by heat treatment, UNG can be used in combination with dUTP toprevent carry-over of amplified products derived from PCR.
This enzyme hydrolyzes uracil from both single- and double-stranded DNA containing dU, but it
not from RNA. It can also be used to prevent carryover of RT-PCR.

[ 2 ] Components
Product name
Uracil-DNA Glycosylase (UNG),
Heat-labile

Conc.

Package

Code No.

Storage

1 U/μL200 U×1

UNG-101－20°C[ 3 ] Properties(1) Unit definition
One unit of UNG is defined as the amount of enzyme required to release 1 nmol uracil fromuracil-containing DNA per hour at 37°C.
(2) Heat inactivation
This enzyme is completely and irreversibly inactivated by incubation for 10 minutes at 50°C.

JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
https://www.toyobo-global.com/seihin/xr/lifescience/tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANHAI) BIOTECH CO., LTD.
Tel(86)-21-58794900.4140
http://www.bio-toyobo.cn/

1https://www.toyobo-global.com/seihin/xr/lifescience/[ 4 ] Source
Produced in E. coli (ung-) strain expressing a recombinant uracil DNA glycosylase gene ofAtlantic cod (Gadus morhua) UNG.

[ 5 ] Buffer composition
Conc.
20 mM
50 mM
1 mM
0.1% (v/v)
50% (v/v)

Component
Tris-HCl, pH 7.5 (25℃)
NaCl
DTT
Tween20Glycerol[ 6 ] Usage example
By adding to PCR reagent and incubating at room temperature (20-25℃) just prior to PCRreaction (RT reaction in RT-PCR), it is possible to eliminate carryover of uracil-containing
amplified products. Since UNG is inactivated during PCR denaturation, there is no effect on thefollowing reaction.
The following table shows a preparation example and reaction cycling conditions using UNG
and our qRT-PCR reagent (THUNDERBIRDTM Probe One-step qRT-PCR Kit, Code: QRZ-101).
Please change the amount of UNG as necessary.
Component

20 μL scale

50 μL scale

RNase free water
2×Reaction Buffer
DNA Polymerase
RT Enzyme Mix
Forward Primer
Reverse Primer
TaqMan® probe
50×ROX Reference dye
Uracil-DNA Glycosylase（UNG）, Heat-labile
RNA solution

X μL
10 μL
0.5 μL0.5 μL10 pmol
10 pmol
4 pmol
0.4 / 0.04 μL
0.4 μL
Y μL

X μL
25 μL
1.25 μL
1.25 μL
25 pmol
25 pmol
10 pmol
1 / 0.1 μL
1.0 μLY μL

Total reaction volume

20 μL

50 μL

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TOYOBO CO., LTD.
Tel(81)-6-6348-3888
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Final conc.
1×0.5 μM
0.5 μM
0.2 μM
1× / 0.1×
0.4 / 1.0 U

CHINA
TOYOBO (SHANHAI) BIOTECH CO., LTD.
Tel(86)-21-58794900.4140
http://www.bio-toyobo.cn/

2https://www.toyobo-global.com/seihin/xr/lifescience/

Cycling conditions
UNG reaction
Reverse transcription
Denaturation
PCR(40～45 cycles)

Denaturation:
Extension (Annealing)

Temperature
25℃
50℃
95℃
95℃
60℃

Time
10 min.
10 min.
1 min.
15 sec.
45 sec.

Package

Code No.

200 U×1

UNG-2010.5 mL×1
1 mL×1
100 reactions

UTP-101
NTP-501
QRZ-101[ 7 ] Related products
Product name
Uracil-DNA Glycosylase (UNG), Heat-labile<Glycerol Free>
dUTP (100 mM)
dNTPs Mixture (A, C, G, U each 2 mM)
THUNDERBIRDTM Probe One-step qRT-PCR Kit

JAPAN
TOYOBO CO., LTD.
Tel(81)-6-6348-3888
https://www.toyobo-global.com/seihin/xr/lifescience/tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANHAI) BIOTECH CO., LTD.
Tel(86)-21-58794900.4140
http://www.bio-toyobo.cn/3