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LABORATORY PROCEDURES FOR PLANT CELL LINES

REFERENCE NO.: PC/1998/2/3


 TITLE: PRELIMINARY QUALITY CONTROL


INTRODUCTION

When a new cell lines arrives for inclusion into the Plant Collection preliminary quality control tests are performed before any further procedure is carried out with the cell line. Only if the cell line is considered to be viable and not contaminated the accessioning procedure is continued by propagation, full quality check and authentication of the culture. The following procedure describes the preliminary quality checks.

PROCEDURE

1. The cell line arrives

1.1. If a frozen cell line is provided a few ampoules are thawed, the rest is stored in liquid nitrogen

1.2. Living callus cultures are examined macroscopically (PC/1998/2/3.1) immediately after arrival. With an aliquot of the callus or suspension material a microscopic examination (PC/1998/2/3.1), a test for fungal and yeast contamination (PC/1998/2/3.2) and a test for bacterial contamination (PC/1998/2/3.3) is carried out.

1.3. If any contamination is detected a letter is sent to the depositor (PC/1998/2/3 Appendix 1) with a request for new material and/or additional information.

1.4. With another aliquot of the callus or suspension material a TTC Test (PC/1998/2/3.4) and a FDA Test (PC/1998//2.3.5) is carried out to check the viability of the culture.

1.5. In case the culture shows low viability a standard letter (PC/1998/2/3 Appendix 2) is sent to inform the depositor and to request more information or a new sample. In case the culture shows no viability the depositor is informed and a new sample requested using standard letter PC/1998/2/3 Appendix 3.

1.6 The remaining cell material of viable cells without contamination is cultivated under appropriate conditions. If a suspension culture gains less than 30 % increase in fresh weight after one week or a callus culture less than 30 % increase in callus volume after four weeks the depositor is informed by a standard letter (PC/1998/2/3 Appendix 4). If no further information or no new culture sample is provided by the depositor, the accession procedure is halted.


Guidelines prepared for CABRI by DSMZ, 20 Jan. 1998
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Copyright CABRI, 1998 

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