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A total of 20 mg of young leaves of each plant and standard nuclear fractions were prepared and mixed.
20 mg of young leaves of rice plantlet were placed at the centre of petri plate and to this, added 2 ml of ice-cold Otto I solution [0.1 M Citric acid, 0.5% (vol/vol) Tween 20].
DNA was extracted from 50 mg of young leaves following the method of Doyle & Doyle [ 36].
Genomic DNA (gDNA) of 71 genotypes was isolated from 50 to 100 mg of young leaves.
Total RNA was extracted from 100 mg of young leaves using the RNeasy Plant Mini Kit (Qiagen).
Genomic DNA of 90 F1 Syrah × Pinot Noir progeny and the two parental genotypes was isolated from 50 100 mg of young leaves.
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For these latter, total genomic DNA extraction was conducted from 150 mg of fresh young leaves, using the DNeasy Plant Mini Kit (QIAGEN, Germany) according to the manufacturer's instructions, with minor modifications: addition of 1% w/v of PVP-40 to the AP1 buffer solution.
Total genomic DNA was extracted from 200 mg of fresh young leaves of the 277 sampled fig trees using the DNeasy Plant Mini Kit (QIAGEN) according to the supplier's instructions with the following modification: 1% of Polyvinylpyrrolidone (PVP 40,000) was added to the buffer AP1.
To determine the ploidy, approximately 20 to 30 mg of fresh young healthy leaf tissue from each sample, in addition to the same amount of internal standard Pisum sativum[ 43], were finely chopped with a blade in a Petri dish containing appropriate buffer [ 44] to lyse the cells and release the nuclei into the suspension.
Approximately 100 mg of tissue from young leaves of a month-old, greenhouse-grown plant was collected and immediately lyophilized.
MG: A significant number of young people are starting and running successful businesses, Mark Zuckerman of Facebook for example, what advice do you have for young, influential, but inexperienced executives like Mark Zuckerman?
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