Convert PCR Primers Into In-Fusion® Primers

 

  • In-Fusion® adaptors, a vector-specific sequences of approximately 15 nr long, will be added on the 5'-end of your primers.
  • Enter (i) sequences of PCR primers 1 and 2, (ii) fragment of vector sequence of about 40 bp with the cloning site in the middle and (iii) definition of the cloning site (i.g. 'CG(A/G)(T/C).CG', dot indicates the position of cut). Make sure that there are at least 15 bp of vector sequence available on each side of the cloning site and that the site is unique.
  • To see how this works, click the EXAMPLE link below to use sequences from the sample.
  • Primer 1 corresponds to the FORWARD ('+' strand) PCR primer. Primer 2 corresponds to the REVERSE ('-' strand) PCR primer.


EXAMPLE. Make In-Fusion® primers for cloning PCR product pcr into the vector vec at the EcoRI restriction site 'GAATTC'.

    ------Primer 1-------    
pcr: 5'- caggattctccgtgaagggataaccagggga- ... ... -gcacgatgaggagtgacactgcctcgggaacct -3'
      ------Primer 2-----  
 
vec: 5'- ...-AAGCTACTTGTTCTTTTTGCACTCGAGAATTCACGCGTGGTACCTCTAGAGTCGACCCGG-... -3'