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From 2012.igem.org

week 12: 16 july - 22 july

Office work

PLRV

This week we designed and ordered all the primers we needed.

Lab work

Biobrick BBa_J04450 was cloned and miniprepped.

HepB

• digestion of the HepB core protein PCR product with pre- and suffix (from 11.July) and ligation into BBa_J04500 (an IPTG inducible promoter with RBS)as well as BBa_PSB1K3.ml (linearized plasmid backbone)
• electro transformation of these constructs with DH5α
• colony PCR (20 samples of the transformants containing HepB core protein + BBa_J04500; 10 samples of the transformants containing HepB core protein + BBa_PSB1K3.ml

-> the ligation and transformation of the HepB core protein into the linear backbone (BBa_PSB1K3.ml) was not successful -> the colony PCR shows that we have 6 out of 20 colonies with the expected insert size (around 600bp)

TuYV

• Mach1 electrocompetent E. coli cells were transformed with last week's ligation mixtures (TuYV coat protein and TuYV coat protein with his-tag). Results were unsatisfying. No colonies were showed on the sample plates.

• 'Coat Protein + readthrough' gene amplicons were digested, ligated into pSB1K3 backbone and transformed into our electrocompetent Mach1 E. coli cells. After two attempts and a colony PCR, results were still unsatisfying.

PLRV

After a two week quest for PLRV infected plant material, we obtained infected potato leafs from the Dutch General Inspection Service for Agricultural seed and seed potatoes.

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