Improvement of an RNA purification method for grapevine (Vitis vinifera L.) suitable for cDNA library construction |
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Authors: | Samia Daldoul Synda Chenenanoui Ahmed Mliki and Michael H?fer |
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Institution: | (1) Laboratoire de Physiologie Moléculaire de la Vigne, B.P.901, 2050 Hammam-Lif, Tunisia;(2) RLP AgroScience GmbH, AlPlanta, Breitenweg 71, 67435 Neustadt an der Weinstrasse, Germany |
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Abstract: | A method is described, which consistently yields high quality total RNA from grapevine. Dissolving of crude RNA pellets in
borate-containing buffer, instead of normally used water before a selective lithium chloride precipitation, was found to be
a critical step, leading to a 2.5-fold increase of yield. The resulting RNA preparations were suitable for standard downstream
applications and also for cDNA library construction. The method worked efficiently and reproducibly and could easily be scaled
from milligram to gram quantities of plant material grown in hydroponic culture, sandy soil or Perlite. It was applied to
different kinds of grapevine tissues (leaves, stem) and, after additional adaptation of the protocol, to roots. |
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Keywords: | cDNA library construction Grape RNA extraction Tris– borate Vitis vinifera L |
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