Numbera | Name | Target/Purpose | Sequence (5' to 3')b |
---|---|---|---|
1 | Bsamut-fwd | Introduction of silent mutation in OspA L10 codon yielding Bsa I site | GGGAATAGGTCT C ATATTAGCCTTAATAGC |
2 | Bsamut-rev | Introduction of silent mutation in OspA L10 codon yielding Bsa I site | TGCTATTAAGGCTAATATG AGACC TATTCC |
3 | Bstmut-fwd | Introduction of silent mutation in mRFP1 V15R16 codons yielding Bst BI site | TGCGCTTCAAGGT T CG A A TGGAGGGCTCCG |
4 | Bstmut-rev | Introduction of silent mutation in mRFP1 V15R16 codons yielding Bst BI site | GGAGCCCTCCAT T CG A A CCTTGAAGCGCATGAAC |
5 | Rmut-oligo | Random mutagenesis oligo | TATTTATTGGGAATAGGTCTC ATATTAGCCTTAATAGCATGTAAGCAAAATGCCTCCTCCNNKNNK GTCATCAAGGAGTTCATGCGCTTCAAGGTTCGAA TGGAGGGCTCCGTG |
6 | Rmut-rev | Generation of double-stranded DNA from Rmut-oligo | CACGGAGCCCTCCATTCGAA CC |
7 | Mutscreen-fwd | Amplification of mutated ospA:mrfp1 region from PflaB | ATGCTATTGCTATTTGCGTTTC |
8 | Mutscreen-rev | Amplification of mutated ospA:mrfp1 region from ospA | ATGGTCTTCTTCTGCATTAC |
9 | Mutscreen-seq | Sequencing of amplified ospA:mrfp1 region from PflaB | AAAGGATTTGCCAAAGTCAG |