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From 2013.igem.org

Contents 1 208 1.1 Main purpose 1.2 Who was in the lab 1.3 Procedure 1.3.1 USER reaction and transformation 1.3.2 Restriction analysis 1.3.3 Primer diluting 1.3.4 PCR to extract Nir from Pseudosomonas with new USER primers 1.4 Results 1.5 Conclusion

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Main purpose

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Who was in the lab

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Henrike, Julia, Gosia, Krystian

Procedure

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USER reaction and transformation

We perform USER reaction, samples are as follows:

• plasmid pZA21 and AMO
• plasmid pZA21 and HAO
• negative control with water instead of insert

Reaction is performed in the same way as on 18-07-2013 with increased amount of insert (up to 14 uL due to low DNA concentration).

Restriction analysis

From last week transformants with AMO and HAO in pZA21 we performed plasmid isolation.

We perform restriction analysis with EcoRI. Expected fragments are as follows:

• For pZA21 with AMO:

- 3309 pz - 2283 pz

• For pZA21 with HAO:

- 2233 pz - 2124 pz - 930 pz

Primer diluting

PCR to extract Nir from Pseudosomonas with new USER primers

Results

Conclusion

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