Lab 208

Main purpose

Make and isolate Nir in 2 fragments and thereafter verify the isolates by amplifying a smaller fragment within the two Nir fragments.

Who was in the lab

Henrike, Kristian

Procedure

Gel electrophoresis

Run 1% agarose gel on overnight PCR products.

Miniprep

Isolate plasmids from samples Sec2, cycAX, TAT 3-1a, TAT 3-2, TAT 2-1 using ethanol precipitation. Actually should have used the Midiprep kit from QIAgen. After ethanol precipitation continued with Midiprep kit step from step 9.

Purification gel

Purification of Nir 2 fragment by gel purification.

Results

Gels

1% agarose gel

• 1kb ladder
• Nir1 high
• Nir1 high
• Nir1 low
• Nir1 low
• broad band ladder
• 43
• 43
• Nir2 GC
• Nir2 GC
• neg for Nir
• AMO
• AMO
• neg for AMO
• 1 kb ladder

Conclusion

The gel verified that Nir1 high is the right fragment but also confirmed that the recurrent fragment on approx. 16 kb from whole Nir isolation is not Nir but unspecific product.

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