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United States Department of Agriculture

Agricultural Research Service

Research Project: MANAGEMENT OF TEMPERATE FRUIT NUT AND SPECIALTY CROP GENETIC RESOURCES Title: Coordinating Cryopreservation Research Projects: Minimizing Lab to Lab Variations

Author
item Reed, Barbara

Submitted to: CryoLetters
Publication Type: Abstract Only
Publication Acceptance Date: August 17, 2009
Publication Date: January 15, 2010
Citation: Reed, B.M. 2010. Coordinating Cryopreservation Research Projects: Minimizing Lab to Lab Variations. International Society for Horticultural Science Meeting. 31(1):77.

Interpretive Summary: The administration of large scale projects that involve multiple laboratories requires careful coordination in order to produce valid data. Due to the variability of laboratory practices it is necessary to have clear and precise protocols and ensure that they are followed by every participant. Examination of each part of the process from plant growth conditions to protocol steps and regrowth parameters was used to determine critical steps that caused most of the variation. Initial planning was vital for successful coordination of the experiments. It was very important for all participants to carefully follow specific written protocols with all details clearly stated. As a result of this experiment we found that our initial protocol was lacking in the detail needed for successful intra-lab coordination. Study of procedures used in each lab allowed us to detail the critical factors that must be addressed in planning a coordinated project. In-vitro plant health is a primary factor. Next operator skills or experience can greatly impact all steps of a protocol so specific training is important. The plant part used must be standardized in any experiments. The physical laboratory conditions play a role as well. Stringent identification and equalization of these critical factors are needed to produce comparable data for the same experiment done in several laboratories.

Technical Abstract: The administration of large scale projects that involve multiple laboratories requires careful coordination in order to produce valid data. Due to the variability of laboratory practices it is necessary to have clear and precise protocols and ensure that they are followed by every participant. Our earlier studies comparing the use of cryopreservation protocols in several laboratories found several critical points that must be controlled so that all laboratories can produce comparable data. Initially the four laboratories following the same protocols had very different results. Examination of each part of the process from plant growth conditions to protocol steps and regrowth parameters was used to determine critical steps that caused most of the variation. Initial planning was vital for successful coordination of the experiments. It was very important for all participants to carefully follow specific written protocols with all details clearly stated. As a result of this experiment we found that our initial protocol was lacking in the detail needed for successful intra-lab coordination. Study of procedures used in each lab allowed us to detail the critical factors that must be addressed in planning a coordinated project. In-vitro plant health is a primary factor. This includes the growth conditions, medium used, length of subculture, hyperhydricity, and any pretreatments. Next operator skills or experience can greatly impact all steps of a protocol from explant excision to manipulation of explants during the procedure, so specific training is important. The plant part used and its location on the plant (apical, axillary or basal) also affects recovery after cryopreservation and this must be standardized in any experiments. The physical laboratory conditions, temperature, humidity and type of laminar air flow, play a role in several of the techniques as well. Stringent identification and equalization of these critical factors are needed to produce comparable data for the same experiment done in several laboratories.

Last Modified: 11/25/2014
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