Primer

Part:BBa_K3103009:Design

Designed by: Devdhi Kasana and Sadie Meunier   Group: iGEM19_IIT_Chicago   (2019-10-16)


pETse_int_N_r


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We did an initial screening with the sequencing primers that flank the gene insertion site in the vector backbone. Every plasmid backbone has a _f and_r sequencing primer which is used to sequencing interred genes, and can also be used to PCR the insert up. When screening by PCR we looked for a specific size (size of insert plus flanking sequence). To confirm OUR gene was inserted, we wanted a specific primer which only amplified our gene of interest. This is a lot more diagnostic. We did this by making primers specific to the gene, and using one of the _f or _r primers in conjunction with the sequencing _r and _f primers.


Source

pCS-PET_VB190412

References