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. 2020 Jul 5;10(13):e3696. doi: 10.21769/BioProtoc.3696
Step Problem Possible solution(s)
7 No SigP in GOI transcript Use the hamster IL2 SigP as described in step 7. See vector pAF92 for additional details (https://www.addgene.org/135929/).
13 GOI segments not in optimal GC% range Order more than one possible primer to find a combination that can amplify the target sequence with the necessary vector overlap segments.