Contents 1 Miniprep for pGFPrrnB with filamentous cell part 1.1 Aims 1.2 Materials and Protocol 1.3 Results 1.4 Discussion 2 Single and Double Digestion of pGFPrrnB with yneA 2.1 Aims 2.2 Materials and Protocol 2.3 Discussion

Miniprep for pGFPrrnB with filamentous cell part

Aims

The aim of this experiment is to obtain stock of the filamentous cell part in pGFP-rrnB for transformation into Bacillus subtilis 168.

Materials and Protocol

Please refer to qiagen minipreps and nanodrop spectrophotometer protocols.

Results

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286.1 µl/ml	304.0 µl/ml	316.3 µl/ml	421.6 µl/ml	518.7 µl/ml	460.1 µl/ml	370.1 µl/ml	377.3 µl/ml	346.0 µl/ml	347.4 µl/ml	202.8 µl/ml	307.4 µl/ml

Table 1: Nanodrop spectrophotometer results. Table represents the amount of plasmid present in µl/ml quantity.

Discussion

The concentration ranged from 286.1 µl/ml to 518.7 µl/ml. Therefore we have obtained high concentration of yneA in pGFPrrnB.

Single and Double Digestion of pGFPrrnB with yneA

Aims

The aim of this experiment is to check if the insert yneA has been inserted into vector pGFPrrnB.

Materials and Protocol

We are doing two digests for pGFPrrnB and yneA:

• Single digest with HindIII;
• Double digest with EcoRI and NheI.

Please refer to restriction digests and gel electrophoresis.

Discussion

Gel electrophoresis will be done on the 25th August, 2010.