Team:Panama/28 September 2010
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==='''September 28'''=== | ==='''September 28'''=== | ||
- | Digestion of | + | 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 | -Electrophoresis |
Revision as of 20:15, 24 October 2010
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
- Table
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