{% extends "layout.html" %} {% block title %}Measurement{% endblock %} {% block lead %}During the three-month-long experimental work, we used a number of experimental measurements, which were:{% endblock %} {% block page_content %}
Competent cells (E. coli DH5α and BL21(DE3)); liquid and solid LB medium; antibiotics.
Liquid LB Formulation
Solid LB Formulation
Plasmid Transformation
Test tubes; liquid LB medium;
Plasmid extraction kit; restriction endonucleases.
Agarose; TAE; EB.
Instruments: Agilent 8890-5977B gas chromatography-mass spectrometer
Chromatographic column: HP-5 MS UI 30 m × 250 μm × 0.25 μm
Injector temperature: 280 ℃
Ionization temperature: 230 ℃
Quadrupole temperature: 150 ℃
Column flow: 1.2 mL/min
Temperature program:
Heating rate (℃/min) | Temperature (℃) | Retention time (min) |
---|---|---|
/ | 60 | 1 |
20 | 200 | 2 |
30 | 300 | 5 |
Transmission line temperature: 280 ℃
Scanning mode: SIM
Quantitative ion: 91
Qualitative ion: 122 65
Standard: 100 μg/mL
KdcA:
10mM Tris-HCl pH=7 140μL
1M MgCl2 10μL
20mM NADH 10μL
100U Adh 10μL
1M Sodium pyruvate 10μL
20mM TPP 10μL
crude enzyme 10μL
Adh1:
10mM Tris-HCl pH=7 160μL
20mM NADH 10μL
10% acetaldehyde 20μL
crude enzyme 10μL
LP (include KdcA and Adh1):
the citric acid / sodium citrate buffer solution pH=6 140μL
1M MgCl2 10μL
20mM NADH 10μL
1M Sodium pyruvate 10μL
20mM TPP 10μL
crude enzyme 20μL