Skip to main content

Advertisement

Table 2 Oligonucleotides used

From: Multiple integration of the gene ganA into the Bacillus subtilis chromosome for enhanced β-galactosidase production using the CRISPR/Cas9 system

Number Sequencea Purpose
S9732N 5′-aaggccaacgaggccAGCAGATGCTGCAGC Construction of pJOE9898.1 flanking sequences
S9733N 5′-aaggccagtctggccGATCAGCTTAATTGCGCTG
S9734N 5′-aaggcctaaatggccGACAGAAGCGGTGTGAC
S9735N 5′-aaggccttattggccGTCGGAGCAATGACGTTTA
S12531 5′-tacgGAATCCCTCACATCTGAAAT Construction of pJOE9898.1 spacer
S12532 5′-aaacATTTCAGATGTGAGGGATTC
S12618 5′-GAAGCACTGATTCTATCACC Colony PCR to prove the deletion by pJOE9898.1
S12619 5′-CATCCAGCCATTCAAATTGA
S9703N 5′-aaggccaacgaggccACTTGAAGGCCGTAACATT Construction of pJOE9899.1, flanking sequences
S9704N 5′-aaggccagtctggccTGCTTAGCGGGCAGT
S9705N 5′-aaggcctaaatggccTGTTAATGATGCAAGGGCT
S9706N 5′-aaggccttattggccCGGCGTCCAATCGTTTTAC
S12533 5′-tacgAGAATTAGAAGATAAAATTG Construction of pJOE9899.1, spacer
S12534 5′-aaacCAATTTTATCTTCTAATTCT
S12620 5′-CAGAGGACTGTACCATGAT Colony PCR to prove the deletion by pJOE9899.1
S12619 5′-CATCCAGCCATTCAAATTGA
S9724N 5′-aaggccaacgaggccCAGGAAGCTGTTGAATCT Construction of pJOE9918.1, flanking sequences
S9725N 5′-aaggccagtctggccGCCCTTCTTTTTCTACTCTA
S9726N 5′-aaggcctaaatggccGCTCATTTTCTTAAAAAGAATATC
S9727N 5′-aaggccttattggccGGCGATAAAGAATTCGAAG
S12529 5′-tacgGCAATCAGCTTCAGTCGAAA Construction of pJOE9918.1, spacer
S12530 5′-aaacTTTCGACTGAAGCTGATTGC
S12660 5′-CGGCCTGTTGAATATTAAAC Colony PCR to prove the deletion by pJOE9918.1
S12619 5′-CATCCAGCCATTCAAATTGA
S9707N 5′-aaggccaacgaggccCATGATGTTTGTCCCTAG Construction of pJOE9957.1, flanking sequences
S9708N 5′-aaggccagtctggccGTTTCATACTGTGCTAAAGA
S9709N 5′-aaggcctaaatggccGTACCCTATAGTTATTTAAATCC
S9710N 5′-aaggccttattggccCCCAAATCGAACACGGTTCA
S12649 5′-tacgATGTCAGCAATACACACAAA Construction of pJOE9957.1, spacer
S12650 5′-aaacTTTGTGTGTATTGCTGACAT
S12715 5′-GAATGCATCCACAGCAGG Colony PCR to prove the deletion by pJOE9957.1
S12716 5′-GGCATAAATCACAGTGGCA
S9795 5′-aaggccaacgaggccGGGCTTGTCTTTATCGTG Construction of pJOE9974.1, flanking sequences
S9796 5′-aaggccagtctggccGATGTGAAGACTGGAG
S9797 5′-aaggcctaaatggccGCCTGGCTTTGATTACGTG
S9798 5′-aaggccttattggccGTGCTCTCCGATAATATGC
S12673 5′-tacgGCTTATATCTATAAACATGA Construction of pJOE9974.1, spacer
S12674 5′-aaacTCATGTTTATAGATATAAGC
S12740 5′-CGGTAAGTCCCGTCTAGC Colony PCR to prove the deletion by pJOE9974.1
S12741 5′-GGGAAGCGTTCACAGTTTC
S12799 5′-aaggccagactggccTAAAAGTACAGTGCCGCTG Amplification of Pgut-ganA
S12800 5′-aaggccatttaggccCCGAAAAGTGCCACCTG
S10494 5′-GACCTCAAAAAGGTCTTTA DNA sequencing of inserts in pJOE8999.1
S11222 5′-CACGCATTGATTGAGTCAG
  1. aSmall letters: nucleotides added for cloning