lab 208

Main purpose

• Gel purification of Cyc and Nir
• USER reaction
• PCR of Nir, His-Tag Sec, His-Tag TAT
• Miniprep re-purification with columns

Who was in the lab

Gosia, Kristian, Natalia, Julia

Procedure

• Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit.
• USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence.
• PCR according to standard PCR protocol.

Samples names:

• 1,2 - Nir 48 (primers 48a, 48b) on template of Nir (too long fragment amplified at the beginning of work with Nir PCR)
• 3,4 - Sec His-Tag, primers 19a, 19b; template: Sec 2
• 5,6 - Tat His-Tag, primers 20a, 20b; template: TAT 2 1
• 7,8 - Tat His-Tag, primers 20a, 20b; template: TAT 3 2
• 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a
• 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag

Results

Conclusion

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