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From 2010.igem.org

Digestion of Rh1ab with Pst1

-Electrophoresis

Clone Rh1ab into the plasmid

-This week

PCR to show the RH1ab gene from within the plasmid it was cloned into

Mutagenesis (Next week)

The gene was purified with the Roche Kit: High Pure PCR Product Purification Kit

The purification of the gene RH1ab was done because it is a requirement in order to be able to use with the pGEM-T Easy Vector system and to clone it into a vector to later do the mutagenesis

Run a gel to see the disgestion of the gene and to verify the cut sites of Pst1

• Table (p.61) and Picture (p.63)

1) We don't see the expected bands, possibly because we aren't using a purified PCR product 2) We can't see the product. We will have to repeat the ligation 3) We observe the band at the expected size

Measuring the concentration of the purification: 96.1 ug/ml

Cloning Protocol of the pGEM-T Vector System

• Table