Abstract
Objective:To study the factors influencing the amplification of sequence-related amplified polymorphism(SRAP) system in Carthamus tinctorius L.and to establish a stable SRAP reaction system,laying a foundation for molecular marker assistant breeding of Carthamus tinctorius L.Methods: Cetyltrimethylammonium bromide method was used to extract the genomic DNA of Carthamus tinctorius L..Twenty-seven tests with 3 factors at 3 levels were designed,the conditions including Taq polymerase concentrations(0.02,0.04,0.06 U/μl),dNTP concentrations(0.15,0.25,0.30 mmol/L)and Primer concentrations(0.15,0.30,0.45 μmol/L);another 8 tests were designed based on the single factor of Mg~(2+) concentrations(0.5,1.0,1.5,2.0,2.5,3.0,3.5 and 4.0 mmol/L).Twenty ng DNA template was added into 25 μl SRAP reaction system;the system was optimized and agarose electrophoresis was used for determination.Results: A SRAP reaction system for Carthamus tinctorius L.was established.In a 25 μl reaction system,Taq polymerase was 0.02 U/μl,dNTP was 0.25 mmol/L,Primer was 0.30 μmol/L,and Mg~(2+) was 3.0 mmol/L.Target bands increased after optimization,with good reproducibility,and the amplification results were satisfactory.Conclusion: The SRAP reaction system in this experiment is suitable for analysis of Carthamus tinctorius L.
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Claims 9
Claim · model-written summary
An SRAP reaction system was established for Carthamus tinctorius L. in a 25 μl reaction volume.
- Taxon
- Carthamus tinctorius L.
Carthamus tinctoriuse0 - Outcome
- SRAP reaction systeme0
- Value
- 25 μle1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e1“In a 25 μl reaction system,Taq polymerase was 0.02 U/μl,dNTP was 0.25 mmol/L,Primer was 0.30 μmol/L,and Mg~(2+) was 3.0 mmol/L”
Claim · model-written summary
The optimal Taq polymerase concentration for the SRAP system is 0.02 U/μl.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse0
- Outcome
- Taq polymerasee1
- Value
- 0.02 U/μle1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e1“In a 25 μl reaction system,Taq polymerase was 0.02 U/μl,dNTP was 0.25 mmol/L,Primer was 0.30 μmol/L,and Mg~(2+) was 3.0 mmol/L”
Claim · model-written summary
The optimal dNTP concentration for the SRAP system is 0.25 mmol/L.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse0
- Outcome
- dNTPe1
- Value
- 0.25 mmol/Le1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e1“In a 25 μl reaction system,Taq polymerase was 0.02 U/μl,dNTP was 0.25 mmol/L,Primer was 0.30 μmol/L,and Mg~(2+) was 3.0 mmol/L”
Claim · model-written summary
The optimal primer concentration for the SRAP system is 0.30 μmol/L.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse0
- Outcome
- Primere1
- Value
- 0.30 μmol/Le1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e1“In a 25 μl reaction system,Taq polymerase was 0.02 U/μl,dNTP was 0.25 mmol/L,Primer was 0.30 μmol/L,and Mg~(2+) was 3.0 mmol/L”
Claim · model-written summary
The optimal Mg2+ concentration for the SRAP system is 3.0 mmol/L.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse0
- Outcome
- Mg~(2+)e1
- Value
- 3.0 mmol/Le1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e1“In a 25 μl reaction system,Taq polymerase was 0.02 U/μl,dNTP was 0.25 mmol/L,Primer was 0.30 μmol/L,and Mg~(2+) was 3.0 mmol/L”
Claim · model-written summary
After optimization, target bands increased in the SRAP amplification.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse0
- Treatment
- optimizatione2
- Outcome
- Target bandse2
- Direction
- increasede2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e2“Target bands increased after optimization,with good reproducibility”
Claim · model-written summary
The optimized SRAP system gave good reproducibility.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse0
- Outcome
- reproducibilitye2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e2“Target bands increased after optimization,with good reproducibility”
Claim · model-written summary
The SRAP amplification results were satisfactory.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse0
- Outcome
- amplification resultse3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A SRAP reaction system for Carthamus tinctorius L.was established”
e3“the amplification results were satisfactory”
Claim · model-written summary
The SRAP reaction system is recommended as suitable for analysis of Carthamus tinctorius L.
- Taxon
- Carthamus tinctorius L.Carthamus tinctoriuse4
- Outcome
- suitable for analysise4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e4“The SRAP reaction system in this experiment is suitable for analysis of Carthamus tinctorius L.”