Guidelines
CELL LINES
PLANT CELL LINES
PLANT VIRUSES
PLASMIDS
PHAGES
MICROORGANISMS
GENOMIC LIBRARIES
PROCEDURES
CATALOGUE
LABORATORY PROCEDURES FOR PLANT CELL LINES
REFERENCE NO.: PC/1998/3/1.4
TITLE: MEASUREMENT OF UV SPECTRA OF UNPURIFIED TOTAL CELL EXTRACTS
INTRODUCTION
Apart from the analysis of extracts by the standard HPLC procedure also UV spectra of the unpurified extracts may be recorded and stored in a computer database.
PROCEDURE
1. The UV spetrophotometer (Perkin-Elmer, UV/VIS spectrometer Lambda 2, 1990) is set to position: on and the computer program PECSS (Vers. 3.2x, 1990) is started.
2. UV light transparent cuvettes (3 ml volume) are washed with methanol (p.A. grade).
3. Standard parameters for the measurement are controlled or set to the standard values.
Standard parameters are:
wavelength: 250 - 500 nm
ord.: A
speed: 480 nm/min
resp.: 1.0 s
smooth: 0 nm
lamps: 3
rec.: 0
assave: Y
aprint: N
4. Methanol is filled in both cuvettes and a "background scan" is performed under standard conditions
5. 2.7 ml methanol + 300 ml of sample are filled in the measuring cuvette and mixed to homogeneity - the control cuvette is left untreated.
6. Before starting the measurement a code name for the sample is entered into the computer.
7. The UV spectrum scan is started - the profile is automatically stored under the given name in the computer database.
8. A printout of the measured UV spectrum is stored.
9. The cuvette is washed two times with ethanol (p.A. grade) and one time with methanol (p.A. grade) before the next extract is filled in.
Guidelines prepared for CABRI by DSMZ, 20 Jan. 1998
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