Cloning – Withering Syndrome Bacteriophage ORFs 20 and 35

Cloned purified PCR products from 20140813 into the pCR2.1 vector using The Original TA Cloning Kit (Life Technologies/Invitrogen), according to the manufacturer’s protocol. Used 2uL of PCR product. Incubated ligation at RT for 15mins. Transformed TOP10 chemically competent cells (Life Technologies/Invitrogen) following the Rapid Transformation protocol (added 4uL of ligation reaction to thawed cells; incubated on ice 5mins; plated 50uL of cells on pre-warmed LB-Amp (50ug/mL) + X-gal (40uL of 40mg/mL stock) plates). Incubated O/N at 37C.


Both cloning reactions look great; lots of potential clones. Will screen nine from each via PCR for detection of our desired insert.

One thought on “Cloning – Withering Syndrome Bacteriophage ORFs 20 and 35

  1. Pingback: Plasmid Isolation – Withering Syndrome Phage pCR2.1/ORF20 and pCR2.1/ORF25 | Sam's Notebook

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