TY  - BOOK
AU  - Barkawi,L.S.
AU  - Tam,Y.-Y.
AU  - Tillman,J.A.
AU  - Normanly,J.
AU  - Cohen,J.D.
TI  - A high-throughput method for the quantitative analysis of auxins
KW  - ACCURACY
KW  - ARTICLE
KW  - AUTOMATION
KW  - CONTROLLED STUDY
KW  - DATA ANALYSIS
KW  - DERIVATIZATION
KW  - HARVESTING
KW  - HIGH THROUGHPUT SCREENING
KW  - HORMONE DETERMINATION
KW  - ISOTOPE DILUTION ASSAY
KW  - MASS FRAGMENTOGRAPHY
KW  - NONHUMAN
KW  - PLANT TISSUE
KW  - PRIORITY JOURNAL
KW  - PROTEIN PROCESSING
KW  - QUANTITATIVE ANALYSIS
KW  - ROBOTICS
KW  - SOLID PHASE EXTRACTION
N2  - Auxin measurements in plants are critical to understanding both auxin signaling and metabolic homeostasis. The most abundant natural auxin is indole-3-acetic acid (IAA). This protocol is for the precise, high-throughput determination of free IAA in plant tissue by isotope dilution analysis using gas chromatography-mass spectrometry (GC-MS). The steps described are as follows: harvesting of plant material; amino and polymethylmethacrylate solid-phase purification followed by derivatization with diazomethane (either manual or robotic); GC-MS analysis; and data analysis. [13 C 6]IAA is the standard used. The amount of tissue required is relatively small (25 mg of fresh weight)and one can process more than 500 samples per week using an automated system. To extract eight samples, this procedure takes ~3 h, whether performed manually or robotically. For processing more than eight samples, robotic extraction becomes substantially more time efficient, saving at least 0.5 h per additional batch of eight samples
UR  - https://drive.google.com/file/d/1LdnqUeHfukFb8Gj-OcxUJ5K7BUnUuUEs/view?usp=drivesdk
ER  -