Team:Panama/28 September 2010

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Cloning Protocol of the pGEM-T Vector System
Cloning Protocol of the pGEM-T Vector System
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Revision as of 11:59, 26 October 2010

iGEM Panama

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September 28

Digestion of Rhlab with Pst1 Since the Rhlab gene is too big we couldn´t complete the sequencing, so we tough that we could test the it with the PstI digest. We check the expected cuts with the NEBcutter (of NEB)

-Electrophoresis

Clone Rh1ab into the plasmid

-This week

PCR to show the RH1ab gene

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

Measurement of the concentration of the purification: 96.1 ug/ml

Cloning Protocol of the pGEM-T Vector System

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