phi29 DNA Polymerase set●This product can amplify genomic DNA from single cell
● Complete removal of contaminating DNA● Suppressing non-speciﬁc ampliﬁcation

A phi29 DNA Polymerase is a replicative polymerase fromphi29.

This polymerase has used Whole Genome Ampliﬁcation（WGA）etc.

F ea t u re s

● Mesophilic enzymes● Extreme strand displacement activity● Extreme processivity● Extreme extensibility● An inherent 3´→5' proofreading exonuclease activity

Example.1： Circular DNA ampliﬁcation by RCA（Rolling Circle Ampliﬁcation）
pUC 18 DNA was ampliﬁed with several kinds of phi29 DNA polymerase. These Amplicons treated withrestriction enzyme (H I andR I) which were detected by agarose gel electrophoresis.（2.7 kbp）

pUC18 copy number（kbp）

M 105 104 103 102

pUC18 copy number
Non-template（kbp）

M 105 104 103 102

pUC18 copy number
Non-template

（kbp）

43
2432

4
3
2

1

1

1

KANTO ProductProduct Ａ

M 105 104 103 102

Non-template

pUC18（2.7 kbp）

Product
B社製品Ｂ● KANTO Ｐｒｏｄｕｃｔ suppresses nonspeciﬁc ampliﬁcation !
Product No.

Product NameStored atSize10195-96

phi29 DNA polymerase set（0.05 mg/mL)-20〜-25℃1set（25 μL × 4）

＊This product contains both phi29 DNA polymerase（0.05 mg/ mL）
25 μL×4 and 10 × Reaction Buﬀer 100 μL×4.
＊This product has been developed through NARO(National Agriculture and Food Research Organization).

Kanto Chemical

Example.2： Human genomic ampliﬁcation by MDA（Multiple Displacement Ampliﬁcation）● Ｎonspeciﬁc binding is not detected

This product used Whole Genome Ampliﬁcation(WGA)
by following protocol. Samples are Human genomic
DNA from human blood or puriﬁed water.

ネガティブコントロールを確認できます鋳型DNA
Template
DNA ( ng )M H

Protocol11ng

1010 ng

0.01
0.1 ng 0.1ng
0.01

Non-1pg
0.001

NTC
templete（kbp）

Template mix
10 × Reaction Buﬀer
100 μM 6R5S primer＊
dDW
Template DNATotal

1.0 μL2.0 μL5.0 μL2.0 μL10.0 μL

23.1
9.4
6.6
4.4
2.3/2.0
0.6

95℃、3 min4℃

Ｅｌｅｃｔｒｏｐｈｏｒｅｓｉｓｂａｎｄｉｎｇｐａｔ
ｔｅｒｎｓ８）＊ 6R5S primer : rN＊rN＊rN＊rN＊rN＊rN＊（6mer）（rN means to mixed base of RNA、＊ means to Phosphorothioated）

10 × Reaction Buﬀer

1.0 μL● This product can amplify DNA of 1 pg than 10 μg

0.1 μL

5,000

25 mM dNTP

0.8 μLdDW

5.8 μL

100 × SYBR GreenⅡ

0.2 μLPPase（1 mg/mL）
1M DTT

0.1 μLphi29Poly（0.05 mg/mL）
Total

2.0 μL10.0 μL

Add to template mix

Fluorescenceintensity
蛍光強度

Reaction mix

M：λⅢ digest marker H：Human genomic DNA
10〜0 : Amplicon after WGA 1 μL

１）Blanco, L. and Salas, M .(1984)., 81, 5325-5329
２）Blanco, L.
. (1989)., 264, 8935-8940
３）Garmendia, C.
. (1992)., 267, 2594-2599
４）Esteban, JA.
. (1993)., 268, 2719-2726
５）Takahashi H,. (2014), 9, 82624
６）Lizardi, PM,. (1998)., 19, 225-232
７）Dean, FB.
. (2002)., 99, 5261-5266８）Akahashi H,. (2009)
47., 609-61

0.01ng

2,000

1pg
1,000

NTC

02468

10

121416

Reactiontime（hr）
反応時間（hr）
Ampliﬁcation plots of human genomic DNA using MDA

20 20

Quantity of ampliﬁed DNA (μg）

References

0.1ng

0DNA合成量（µg）

4℃1ng

3,00030℃、16 hr（measuring the ﬂuorescence every 10 min）
65℃、10 min

10ng

4,000

16 1612 1288
4400

10 ng

101 ng0.01 ng

0.1 ng1 pg

NTC

1 0.1 0.01 0.001 NTC
Quantity of template DNA
鋳型 DNA（ng）

Quantity of ampliﬁed DNA on ampliﬁcation

KANTO CHEMICAL CO., INC.
REAGENT DIVISION

East Muromachi Mitsui BLDG, 2-1, Nihonbashi Muromachi 2-chome,
Chuo-ku, Tokyo, 103-0022, JAPAN
Telephone+81-3-6214-1092
Telefax+81-3-3241-1053
http://www.kanto.co.jp
E-mail; kanto-61@gms.kanto.co.jp
2015. November.