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袁存权等
, 2011,
刺槐
SRAP-PCR
反应体系优化及引物筛选
,
分子植物育种
Vol.9 No.25 (doi: 10.5376/mpb.cn.2011.09.0025)
1186
表
1 SRAP
引物序列信息
Table 1 The primer sequences used in SRAP analysis
编号
Serial No.
正向引物
(5'
-
3')
Forward primers (5'
-
3')
编号
Serial No.
反向引物
(5'
-
3')
Reverse primers (5'
-
3')
Me1
TGA GTC CAAACC GGA TA
Em1
GAC TGC GTA CGAATT AAT
Me2
TGA GTC CAAACC GGA GC
Em2
GAC TGC GTA CGAATT TGC
Me3
TGA GTC CAAACC GGAAT
Em3
GAC TGC GTA CGAATT GAC
Me4
TGA GTC CAAACC GGA CC
Em4
GAC TGC GTA CGAATT TGA
Me5
TGA GTC CAAACC GGAAG
Em5
GAC TGC GTA CGAATT AAC
Me6
TGA GTC CAAACC GGA CA
Em6
GAC TGC GTA CGAATT GCA
Me7
TGA GTC CAAACC GGA CG
Em7
GAC TGC GTA CGAATT CAA
Me8
TGA GTC CAAACC GGA CT
Em8
GAC TGC GTA CGAATT CAC
Me9
TGA GTC CAAACC GGA GG
Em9
GAC TGC GTA CGAATT CAG
Me10
TGA GTC CAAACC GGAAA
Em10
GAC TGC GTA CGAATT CAT
Me11
TGA GTC CAAACC GGAAC
Em11
GAC TGC GTA CGAATT CTA
Me12
TGA GTC CAAACC GGA GA
Em12
GAC TGC GTA CGAATT CTC
Me13
TGA GTC CAAACC GGAAG
Em13
GAC TGC GTA CGAATT CTG
表
2 SRAP-PCR
优化反应中的各因素水平
Table 2 Each factors and levels of SRAP-PCR optimization reaction
水平
镁离子
(mmol/L)
脱氧核苷酸
(mmol/L)
Taq
聚合酶
(U/25 µL)
引物
(µmol/L)
DNA
模板
(ng/25 µL)
Level Mg
2+
(mmol/L)
dNTPs (mmol/L)
Taq
DNA polymerase (U/25 µL) Primer (µmol/L) DNA template (ng/25 µL)
1
1.5
0.1
0.5
0.1
10.0
2
2.0
0.2
1.0
0.2
30.0
3
2.5
0.3
1.5
0.3
50.0
4
3.0
0.4
2.0
0.4
70.0
表
3 SRAP-PCR
反应中各因素水平的
L
16
(4
5
)
正交试验设计
Table 3 L
16
(4
5
) orthogonal design of each factors and levels of SRAP-PCR reaction
编号
Serial No.
镁离子
(mmol/L)
Mg
2+
(mmol/L)
脱氧核苷酸
(mmol/L)
dNTPs (mmol/L)
Taq
聚合酶
(U/25µL)
Taq
DNA polymerase (U/25µL)
引物
(µmol/L)
Primer (µmol/L)
DNA
模板
(ng/25 µL)
DNA template (ng/25 µL)
1
1.5
0.1
0.5
0.1
10.0
2
1.5
0.2
1.0
0.2
30.0
3
1.5
0.3
1.5
0.3
50.0
4
1.5
0.4
2.0
0.4
70.0
5
2.0
0.1
1.0
0.3
70.0
6
2.0
0.2
0.5
0.4
50.0
7
2.0
0.3
2.0
0.1
30.0
8
2.0
0.4
1.5
0.2
10.0
9
2.5
0.1
1.5
0.4
30.0
10
2.5
0.2
2.0
0.3
10.0
11
2.5
0.3
0.5
0.2
70.0
12
2.5
0.4
1.0
0.1
50.0
13
3.0
0.1
2.0
0.2
50.0
14
3.0
0.2
1.5
0.1
70.0
15
3.0
0.3
1.0
0.4
10.0
16
3.0
0.4
0.5
0.3
30.0