Via gDNA analysis and PCR was the specific area of the rsh gene extracted and amplified where the nucleotide substitution: C to T (Folkers et al., 2006 ) should be for the rsh1 mutation. The amplicon was Sanger sequenced which proved the nucleotide substitution.
1% Agarose gel with 100bp marker and PCR1 rsh , PCR2 rsh and PCR3 rsh or PCR1 rut , PCR2 rut and PCR3 rut , respectively. The template pCFD6 was used with a concentration of 640 pg/µl.
1% Agarose gel with 100bp marker and PCR1 rsh , PCR2 rsh and PCR3 rsh or PCR1 rut , PCR2 rut and PCR3 rut , respectively. The template pCFD6 was used with a concentration of 64 pg/µl (1:10 dilution).
1% Agarose gel with 100bp marker and PCR1 rsh , PCR2 rsh and PCR3 rsh or PCR1 rut , PCR2 rut and PCR3 rut , respectively. The template pCFD6 was used with a concentration of 64 pg/µl (1:10 dilution). 50µl of 5xQ5 High GC Enhancer was added to the PCR mix.
1% Agarose gel with 100bp marker and PCR1 rsh , PCR1 rut . The template pCFD6 was used with a concentration of 128 pg/µl (1:5 dilution). 50µl of 5xQ5 High GC Enhancer was added to the PCR mix.
1% Agarose gel with 100bp marker and PCR1 rsh , PCR2 rsh and PCR3 rsh or PCR1 rut , PCR2 rut and PCR3 rut , respectively. The template pCFD6 was used with a concentration of 640 pg/µl. The Phusion DNA Polymerase was used instead of the Q5 High-Fidelity DNA Polymerase.