Primers for QRT-PCR | |||
---|---|---|---|
Name | Purpose | Location | Sequence (5' to 3') |
MA0326 | Â | spacer #2 | CCTGGTTGAGGTTAGCGTTGA |
PCR of spacer #1 | Â | Â | Â |
MA0327 | Â | outside CRISPR | AATTCGGTGGCCAGTTGTTC |
MA0328 | Â | sense strand | CAGGAAGCCACCAAGGAT |
PCR of hisS Pcar_1041 | Â | Â | Â |
MA0329 | Â | antisense strand | TGGGAGCCGAGTTGATTG |
MA0441 | Â | sense strand | CAAACTGATTGCCGTTCCTT |
PCR of hisZ GSU3307 | Â | Â | Â |
MA0442 | Â | antisense strand | AGGCCGATGAGTTCTACGC |
Primers for construction of hisS transgenic strain MA159 | |||
Name | Purpose | Sequence (5' to 3') | |
MA0330 | Â | TGACATCTCGCTGGACCGGG | |
PCR on 5' side of hisS GSU1659 | Â | Â | Â |
MA0331 | Â | CTATGCTAGCACTAGTTTGTAATCATGAACGTACCTACTC CTTTAATTG | |
MA0332 | Â | GTACGTTCATGATTACAAACTAGTGCTAGCATAGCAATAC CTGCATTG | |
PCR on 3' side of hisS GSU1659 | Â | Â | Â |
MA0333 | Â | AGTCCATTCCTCCTGTGG | |
MA0334 | Â | AAGGGATCTATCATGAGCATATCAGGCATTAAGGG | |
PCR of hisS Pcar_1041 | Â | Â | Â |
MA0335 | Â | GCGCGGCGCGACTAGTTTCCTCGTGTCTTTTCC | |
MA0052 | Â | TGCATATGGCTCTAGAATAACTTCGTATAGC | |
gentamicin marker | Â | Â | Â |
MA0053 | Â | TCGATAAGCTTCTAGAATAACTTCGTATAATG | |
Oligonucleotides for construction of chimeric CRISPR expression plasmid pMA35 | |||
Name | Purpose | Sequence (5' to 3') | |
MA0269 | Â | ACATGTCACTGCCCGCTTTCCAGTC | |
PCR of lacI- taclacUV5 promoter | Â | Â | Â |
MA0270 | Â | GCATGCGTGTGAAATTGTTATCCGC | |
MA0429 | Â | AATTCGGTTCATCCCCGCGCATGCGGGGAACACATACAT GAGGGCAAACGCCTTTTGGCCGGCGGCGGTTCATCCCCG CGCATGCGGGGAACACG | |
synthetic CRISPR of spacer #1 | Â | Â | Â |
MA0430 | Â | GATCCGTGTTCCCCGCATGCGCGGGGATGAACCGCCGCC GGCCAAAAGGCGTTTGCCCTCATGTATGTGTTCCCCGCAT GCGCGGGGATGAACCG | |
MA36R | sequencing | CGACATCATAACGGTTC |