PCR: Mr T-Less (MrT)
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ID:  104
Amplifluor: No
Primers:
Nucleotide sequence (5´ ⇒ 3´)
Primer name Sequence
MrT-F CCGCAATGTTCTACTGGTCA
MrT RWT GAAAGTTGATGCACTCTGGCGCGTATCA
MrT-RM CACTGCGGCTGGAGAACTTCCTCAAGTTGATGCACTCTGGCGCGTGCCG
Reagents:
Reagents / Constituents Commercial name Stock
Concentration
Volume (μL)
DNA Sample NA ng/μl 1.0
10x Buffer 20.0 mM MgCl2 5.0
dNTPs 10.0 mM 0.5
20x amplifluor SNP FAM Primer NA μM NA
20x amplifluor SNP JOE Primer NA μM NA
Primer:  MrT-F 10.0 μM 2.0
Primer:  MrT RWT 10.0 μM 4.0
Primer:  MrT-RM 10.0 μM 2.0
Amplifluor Primer Volume NA
Water 32.85
Taq Polyermase 0.15
25mM MgCl2 2.0
Strategy:
Steps Temp (°C ) Time (seconds)
Hot start: false    
Initiation/Melting 94.0 120
Denaturation 94.0 20
Annealing 65.0 20
Elongation 72.0 20
Number of cycles
(repeat denature, anneal & elongate):
35
   
Strand completion (i.e. 72°C, 10 min) 72.0 180
Finish (i.e. 4°C, indefinite) 10.0 Hold
Electrophoresis:
% Agarose 4.5
Volt 400.0
Estimated Running Time (min) 20.0
Number Band (bp) Genotype
1. 151 WT/WT
2. 172 MrT/MrT
Build: 2024.03.19 Non-profit supported by the Australian government  |  © Copyright 2024 Australian Phenomics Facility Last updated: 16/05/2024