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

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

Materials and Protocol

Please refer to PCR purification.

Restriction Digest

Aim

To digest the PCR products of pSB1C3 and yneA from PCR purification with enzymes EcoR1 and Nhe1.

Materials and Protocol

Please refer to restriction digest.

Results, Discussion and Conclusion

We run the digested products with gel electrophoresis to determine whether the digest worked.

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 gel did not show any band when we looked at it from GelDoc.

Conclusion

The restriction digest did not work, so we will repeat the protocol again tomorrow.