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From 2010.igem.org

Contents 1 yneA 1.1 PCR (Repeat) 1.1.1 Aim 1.1.2 Materials and Protocol 1.2 PCR Purification 1.2.1 Aim 1.3 PCR Purification 1.3.1 Aim 1.3.2 Materials and Protocol 1.4 Digestion 1.4.1 Aim 1.4.2 Materials and Protocol 1.5 Gel extraction 1.5.1 Aim 1.5.2 Materials and Protocol 1.5.3 Results 1.5.4 Conclusion

yneA

PCR (Repeat)

Aim

To repeat the PCR that we did yesterday using the correct rocF primers.

Materials and Protocol

Please refer to PCR.

PCR Purification

Aim

PCR Purification

Aim

To remove unwanted primers, taq polymerase, buffer and salts to obtain pure DNA.

Materials and Protocol

Please refer to PCR purification.

Digestion

Aim

To digest the PCR products of pSB1C3 and yneA from PCR purification.

Materials and Protocol

Please refer to restriction digest.

Gel extraction

Aim

To purify the DNA of yneA and pSB1C3 by extracting the bands from the gel after running gel electrophoresis. Concentration of DNA is then checked with NanoDrop.

Materials and Protocol

Please refer to:

• gel electrophoresis,
• gel extraction and
• nanodrop spectrophotometer.

Results

The bands we got from gel electrophoresis is very faint.

Conclusion

We realized that we used the wrong rocF primer, so we repeat the whole procedure from PCR again.