TBX1 genotyping
The following are the sequences of the primers used to genotype Tbx1:
TBX1For:
5 TTG GTG ACG ATC ATC TCG GT 3
Tbx1Rev:
5 ATG ATC TCC GCC GTG TCT AG 3
Mut:
5 AGG TCC CTC GAA GAG GTT CA 3
PCR Program to genotype TBX1
Touch Down Protocol:
Step1: 94 1min
Step2: 94 1min
Step3: 68 1min
Step4: 72 1min
Step5: 2 times to Step 2
Step6: 94 1min
Step7: 66 1min
Step8: 72 1min
Step9: 2 times to Step 6
Step10: 94 1min
Step11: 64 1min
Step12: 72 1min
Step13: 2 times to Step 10
Step14: 94 1min
Step15: 62 1min
Step16: 72 1min
Step17: 30 times to Step 14
Step18: 72 - 7 min
Step19: 4 24:00:00
Step20: End
OR
Step1: 94 3min
Step2: 94 1min
Step3: 58 45sec
Step4: 72 45sec
Step5: 35 times to Step 2
Step6: 72 7min
Step7: 4 24:00:00
Step8: End
PCR Reaction:
Primer1(For) 1
Primer2(Rev) - 0.5
Primer3(Mut) 0.5
QSolution 10
10XPCRBuffer 5
Taq 0.5
DNA 1
Water - 31.5
Wildtype band : 350bp
Mutant Band : 450bp
Het: 350bp and 450bp
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