lab 208

Main purpose

Who was in the lab

Kristian, Henrike, Gosia

Procedure

PCR (promoter library)

PCR to amplify USER fragments of the constitutive SPL, the constitutive reference promoter and the arabinose reference promoter.

Used 5% DMSO, 2 uL of mM MgCl2 and GC-buffer.

constitutive SPL

• template: pZA21::RFP
• primers: 52a, 52b1

constitutive reference

• template: pZA21::RFP
• primers: 52a, 52b2

arabinose reference

• template:
• primers: 51a,51b1

program

Results

Gel of SLP screening PCR products (yesterday)

Each gel is representing one row; from left to right and up to down is row A to H:

Conclusion

SLP screening PCR

The PCR run with condition E (5% DMSO, 2mM MgCl2 with GC buffer) had the highest flexibility; it yields the expected product at different annealing temperatures: 58.4 C, 59.6, C 60.6 C, 61.9 C, 63.2 C, 64.6 C, 65 C.

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