The oligonucleotide is not validThe oligonucleotide is not validThe oligonucleotide is not validThe oligonucleotide is not valid RF Cloning Project

Restriction Free Cloning

Construct Name
pPAGFP-C1-https://bitly.cx/6jHom

Forward Primer
Plasmid annealing = 0°C     Target annealing = 0°C    Length = 60
XXXTTXXXXTXXCXXXXXXTTXXXXTXXCXXXXXXXTTXXXXTXXCXXXXXXTTXXXXTX


Reverse Primer
Plasmid annealing = 0°C     Target annealing = 0°C    Length = 60
NNAANNNNNNGNNANNNNAANNNNNNGNNANNNNANNNNNGNNANNNNAANNNNNNGNNA

1° PCR Size New Plasmid Size Insert Sites Insert Size
358bps 575bps
1-2
288bps

2° PCR conditions
Extension Time ng of insert ng of plasmid
0:11 mins 85.9 3.5

New construct
ATGC = Original plasmid sequence. ATGC = Inserted sequence.

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Feb 06 2026



Plasmid markers and restriction enzymes:







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