Selected article for: "PCR reaction and second PCR reaction"
Author: Václav Vopálenský; Michal Sýkora; Tomáš Mašek; Martin Pospíšek
Title: Messenger RNAs transcribed from yeast linear cytoplasmic plasmids possess unconventional 5’ and 3’ UTRs and suggest a novel mechanism of translation
  • Document date: 2018_5_17
    • ID: foskvkwn_24
    Snippet: Both primary PCR products, which had defined overlapping ends, were synthesized as follows: 5 min at 95°C; then 30 cycles of 30 sec at 94°C, 30 sec at 56.2°C, and 90 sec at 72°C; and finally, 10 min at 72°C. Then, 1 µl of each PCR mixture was used without purification for a second PCR reaction using Pfu DNA polymerase (Fermentas), the primers ORF3_SAM_del_F1 and ORF3_SAMdel_R2 and following conditions: 5 min at 95°C; then 35 cycles of 30 s.....
    Document: Both primary PCR products, which had defined overlapping ends, were synthesized as follows: 5 min at 95°C; then 30 cycles of 30 sec at 94°C, 30 sec at 56.2°C, and 90 sec at 72°C; and finally, 10 min at 72°C. Then, 1 µl of each PCR mixture was used without purification for a second PCR reaction using Pfu DNA polymerase (Fermentas), the primers ORF3_SAM_del_F1 and ORF3_SAMdel_R2 and following conditions: 5 min at 95°C; then 35 cycles of 30 sec at 94°C, 30 sec at 60.1°C, and 2 min at 72°C; and finally, 10 min at 72°C).

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