Gel extraction of amplified plasmid pSB1C3 for rocF

Aim

The aim of this experiment is to perform gel extraction of the band containing the amplified fragment of the plasmid vector pSB1C3.

Materials and Protocol

Please refer to: Gel extraction.

Result

For plasmid vector pSB1C3, we used 1.5% agarose gel and run the gel on 80 Volts for better resolution. We used 1Kb DNA ladder (Promega) for the gel containing plasmid vector.

Table 1: Table represents the size of the plasmid vector pSB1C3 represented as bands on the gel.

1. Weight of gel 0.3g, nanodrop value 27.3ng/microL
2. Weight of gel 0.4g, nanodrop value 21.6ng/microL

Discussion

We found a band of the appropriate size and cut it out and placed the part of the gel in a 2ml eppendorf tube.

Conclusion

Gel electrophoresis was successful and we did gel extraction of the fragment containing plasmid pSB1C3.

Natto Rehydration

Aims

We aimed to rehydrate and plate the natto strain (Bacillus subtilis ATCC 31578) bought from DSMZ, Germany. We used Penassay G- Thy Medium to rehydrate the cells.

Materials

• Pennassay G- Thy medium:

1. Pennassay broth- 17.5g
2. Glucose- 20g
3. Thymine- 0.05g
4. Distilled water- 1000ml

• Pennassay broth(per l of water):

1. peptone 10g
2. Yeast extract 1.5g
3. Sodium chloride 3.5g
4. Glucose 1.0g
5. Potassium phosphate dibasic 3.68g
6. Potassium phospahte monobasic 1.32g
7. Beef extract 1.5g

ph7 store at 2-8oC