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Blood, Vol. 111, Issue 5, 2606-2614, March 1, 2008
A dyskerin motif reactivates telomerase activity in X-linked dyskeratosis congenita and in telomerase-deficient human cells
Blood Machado-Pinilla et al.
111: 2606
Supplemented materials for: Machado-Pinilla et al
Primer sets used to amplify specific sequences were:
5′-CGG AAG AGT GTC TGG AGC AA-3′ and 5′-GGA TGA AGC GGA GTC GGA-3′ for hTERT; 5′-TCT AAC CCT AAC TGA GAA GGG CGT AG-3′ and 5′-GTT TGC TCT AGA ATG AAC GGG GAA G-3′ for hTR; 5′-ATG GCG GAT GCG GAA GTA ATT-3′ and 5′-CCC CTT CAA TAG CAT TGT GC-3′ for DKC; 5′-CCA GCA GCG ACT CTG AGG-3′ and 5′-CCA AGA CGT TGT GTG TTC-3′ for human c-MYC; 5′-CTG TTT GAA GGC TGG ATT T-3′ and 5′-TAC AGG CTG GAG GTG GAG CA-3′ for Mouse c-MYC; 5′-GTG TAA AAC GCA GCT CAG TAA CAG TCC G-3′ and 5′-GGT ATG GAA TCC TGT GGC ATC CAT GAA A-3′  -actin mouse. The PCR conditions were: for hTERT 94°C 45s, 60°C 45s, 72°C 90s, for 20 cycles; for hTR 94°C 45s, 55°C 45s, 72°C 90s, for 20 cycles. The conditions for DKC were: 94°C 40s, 60°C 60s, 72°C 120s, for 20 cycles. The conditions for human c-MYC were: 94°C 30s, 60°C 60s, 72°C 60s, for 25 cycles. The conditions of GAPDH were 94°C 60s, 60°C 60s, 72°C 120s, for 35 cycles. The conditions for mouse c-MYC were: 94°C, 60s, 60°C 60s, 72°C, 60s, for 25 cycles. The conditions for mouse -actin were: 94°C, 60s, 60°C, 60s, 72°C 60s, for 25 cycles. Primers for band shift assays were:
TEL1 5′TAA CCC TAA CCC TAA GCG AAT TCG TCA TGC GAA TTC GCT TAG GGT TAG GGT TAG GGT TAG GGT TAG GGT TAG GGT TAG GG-3′, CXext: 5′GTG CCT TAC CCT CTA CCC TTA CCC TAA-3′
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