scholarly journals An improved method for the rapid isolation of RNA from Arabidopsis and seeds of other species high in polyphenols and polysaccharides

2018 ◽  
Vol 28 (4) ◽  
pp. 360-364
Author(s):  
Steven Footitt ◽  
Sajjad Awan ◽  
William E. Finch-Savage
Keyword(s):  
Polyethylene Glycol ◽  
Brassica Oleracea ◽  
Rna Extraction ◽  
Improved Method ◽  
Sinapis Arvensis ◽  
Rapid Isolation ◽  
Arabidopsis Seed ◽  
Gelatinous Matrix ◽  
Field Soils ◽  
Carry Over

AbstractSeeds are notoriously high in polyphenols and polysaccharides, which reduce RNA quality and yield, and interfere with downstream applications. We present simple modifications to a rapid RNA extraction protocol for use with seeds. The inclusion of polyethylene glycol in place of polyvinylpyrrolidone reduced polyphenol and polysaccharide contamination. In addition, replacing NaCl with KCl improved the RNA yield from Arabidopsis seeds still bound by mucilage. On extraction of Arabidopsis seed recovered from field soils clean RNA pellets with no accompanying gelatinous matrix (polysaccharide) were seen, with A260/230 ratios greater than 1.8 confirming the lack of polysaccharide carry-over. WhenBrassica oleraceaandSinapis arvensisseeds were extracted, 260/230 ratios greater than 1.8 were seen. RNA yields in excess of 10 µg per 100 mg seed suitable for RT-QPCR were obtained.

scholarly journals An improved method for RNA extraction from common bean seeds and validation of reference genes for qPCR

2017 ◽  
Vol 17 (2) ◽  
pp. 150-158 ◽  
Author(s):  
Wendell Jacinto Pereira ◽  
Priscila Zaczuk Bassinello ◽  
Claudio Brondani ◽  
Rosana Pereira Vianello
Keyword(s):  
Common Bean ◽  
Reference Genes ◽  
Rna Extraction ◽  
Improved Method

scholarly journals RT-LAMP has high accuracy for detecting SARS-CoV-2 in saliva and naso/oropharyngeal swabs from asymptomatic and symptomatic individuals

2021 ◽  
Author(s):  
Stephen P Kidd ◽  
Dan Burns ◽  
Bryony Armson ◽  
Andrew D Beggs ◽  
Emma L. A Howson ◽  
...  
Keyword(s):  
Preparation Method ◽  
Rna Extraction ◽  
Improved Method ◽  
Diagnostic Specificity ◽  
Sample Preparation Method ◽  
Viral Loads ◽  
Healthcare Settings ◽  
Frequent Interval ◽  
Oropharyngeal Swab ◽  
Asymptomatic Individuals

Previous studies have described RT-LAMP methodology for the rapid detection of SARS-CoV-2 in nasopharyngeal/oropharyngeal swab and saliva samples. Here we describe the validation of an improved simple sample preparation method for Direct SARS-CoV-2 RT-LAMP, removing the need for RNA extraction, using 559 swabs and 86,760 saliva samples from asymptomatic and symptomatic individuals across multiple healthcare settings. Using this improved method we report a diagnostic sensitivity (DSe) of 70.35% (95% CI 63.48-76.60%) on swabs and 84.62% (79.50-88.88%) on saliva, with diagnostic specificity (DSp) 100% (98.98-100.00%) on swabs and 100% (99.72-100.00%) on saliva when compared to RT-qPCR. Analysing samples with RT-qPCR ORF1ab CT values of <25 and <33 (high and medium-high viral loads, respectively), we found DSe of 100% (96.34-100%) and 77.78% (70.99-83.62%) for swabs, and 99.01% (94.61-99.97%) and 87.32% (80.71-92.31%) for saliva. We also describe RNA RT-LAMP (on extracted RNA) performed on 12,619 swabs and 12,521 saliva samples to provide updated performance data with DSe and DSp of 95.98% (92.74-98.06%) and 99.99% (99.95-100%) for swabs, and 80.65% (73.54-86.54%) and 99.99% (99.95-100%) for saliva, respectively. We also report on daily samples collected from one individual from symptom onset where both Direct and RNA RT-LAMP detected SARS-CoV-2 in saliva collected on all six days where symptoms were recorded, with RNA RT-LAMP detecting SARS-CoV-2 for an additional further day. The findings from these studies demonstrate that RT-LAMP testing of swabs and saliva is potentially applicable to a variety of use-cases, including frequent, interval-based testing of saliva from asymptomatic individuals via Direct RT-LAMP that may be missed using symptomatic testing alone.

scholarly journals Novel method for the rapid isolation of RPE cells specifically for RNA extraction and analysis

2012 ◽  
Vol 102 ◽  
pp. 1-9 ◽  
Author(s):  
Cynthia Xin-Zhao Wang ◽  
Kaiyan Zhang ◽  
Bogale Aredo ◽  
Hua Lu ◽  
Rafael L. Ufret-Vincenty
Keyword(s):  
Rna Extraction ◽  
Rpe Cells ◽  
Rapid Isolation ◽  
Novel Method

Yield and injury effects on vegetable crops planted in flumetsulam-treated soil

1999 ◽  
Vol 79 (3) ◽  
pp. 417-420 ◽  
Author(s):  
J. O'Sullivan ◽  
R. J. Thomas ◽  
W. J. Bouw
Keyword(s):  
Solanum Tuberosum ◽  
Brassica Oleracea ◽  
Solanum Tuberosum L ◽  
Low Ph ◽  
Yield Reduction ◽  
Vegetable Crops ◽  
Treated Soil ◽  
Crop Tolerance ◽  
Ph Values ◽  
Carry Over

The effect of flumetsulam on several vegetable crops grown following soybean was investigated over a 4-yr period. Flumetsulam was applied preplant incorporated to soybean at 0, 70 and 140 g a.i. ha−1 from 1992 to 1994. In trials 1 yr after flumetsulam application, cabbage (Brassica oleracea var. capitata), potato (Solanum tuberosum L.), tomato (Lycopersicon esculentum L.), and pepper (Capsicum annuum L.), showed visual injury symptoms and reduced yields in 1993, while only cabbage showed visual injury symptoms and reduced yields in 1994 and 1995. Cucumber (Cucumis sativus) showed injury symptoms only in 1993 and had no yield reduction. Cabbage also showed visual injury and reduced yields 2 and 3 yr following a 1992 flumetsulam application. Broccoli (Brassica oleracea var. italica) and cauliflower (Brassica oleracea var. botrytis) showed visual injury symptoms in 1995, 3 yr after flumetsulam application. Generally, crop tolerance decreased linearly as application dose increased from 0 to 140 g ha−1. Minor visual injury symptoms did not correspond with a reduced yield. Injury was most severe on sites with the lowest soil pH values. Based on these studies, effects of flumetsulam from normal use rates (70 g ha–1) will carry over to injure-sensitive crops like cabbage, in the rotation for up to 3 yr, especially in low pH soils (pH 6.1 or less). Otherwise, a 22-mo planting interval is adequate to protect other vegetable crops in the rotation from injury and yield reductions. Key words: Crop injury, herbicide carryover, flumetsulam, yield

scholarly journals 836 PB 352 EFFECTS OF PRIMING ON MUSKMELON SEED STORABILITY

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 553a-553
Author(s):  
Melkizedek O. Oluoch ◽  
Gregory E. Welbaum
Keyword(s):  
Polyethylene Glycol ◽  
Moisture Content ◽  
Water Potential ◽  
Cucumis Melo ◽  
Seed Storage ◽  
Germination Percentage ◽  
Saturated Soils ◽  
Field Soils ◽  
Controlled Deterioration ◽  
Seed Storability

Priming (controlled hydration followed by drying) has been shown to decrease seed storage life in some species The germinablity of primed (0.3 M KNO3, 6 d, 25°C) and unprimed muskmelon (Cucumis melo L., cv. PMR 45) seeds were compared after storage for 9 yrs at less than 20°C and 6% moisture content (MC) (dwt basis). Germination performance was compared at 30°C in water and polyethylene glycol solutions of -0.2, -0.4, -0.6, -0.8, and -1.0 MPa water potential or in water at 15, 20, and 25°C. Seeds were also germinated in field soils at 17, 19, and 21% (dwt. basis) MC in a greenhouse, Some seeds were subjected to controlled deterioration at 20% MC and 45°C for 72 hrs prior to testing. The germination percentage and rate of stored, primed seeds at 30°C and all water potentials was less than stored, unprimed seeds. At 30°C, stored, unprimed seeds germinated more rapidly and to higher percentages at -0.2 MPa than in water, while germination percentages and rates of stored, primed seeds were essentially the same. At 15, 24, and 25°C, stored, primed seeds outperformed unprimed seeds in all germination tests. In saturated soils at 21% MC, there was no germination of either stored, primed or unprimed seed. At 17% soil MC, stored, primed seeds germinated 73% compared to only 56% for unprimed seeds. The enhancement due to priming was retained after 9 yrs of storage at germination temperatures <30°C. At higher temperatures, the germination of unprimed seeds was superior to primed.

scholarly journals Does Primed Seed Improve Stand Establishment and Yield of Broccoli?

1995 ◽  
Vol 5 (4) ◽  
pp. 314-317 ◽  
Author(s):  
Lewis W. Jett ◽  
Gregory E. Welbaum ◽  
Charles R. O'Dell ◽  
Ronald D. Morse
Keyword(s):  
Molecular Weight ◽  
Polyethylene Glycol ◽  
Brassica Oleracea ◽  
Seedling Emergence ◽  
Plant Competition ◽  
Seed Priming ◽  
Stand Establishment ◽  
The Third ◽  
The Mean ◽  
Mean Time

The effect of matric and osmotic seed priming on stand establishment and maturity of broccoli (Brassica oleracea L. var. italica) was investigated in three years at two locations in Virginia. Seeds (`Earlidawn') were primed at 1.1 MPa (68F for 7 days) either osmotically in polyethylene glycol (8000 molecular weight) or metrically in vermiculite (horticultural grade no. 2). In the frost year of the study, seeds were hand-seeded in August into crustprone soil with a mean temperature of 82F, and there were no differences in the percentage or mean time of seedling emergence between osmotic- and matric-primed seeds. Under cooler temperatures during the remaining two years of the study, priming increased the percent emergence and decreased the mean time of emergence by about 15 hours. Primed seeds did not increase yields or accelerate maturity in two out of three years. In the third year, the spread of seedling emergence times was less for primed seeds, which reduced plant-to-plant competition and hastened maturity. The primary benefit of primed broccoli seeds was faster emergence, which increased stands by reducing exposure to stresses that decrease emergence.

scholarly journals An improved method for the rapid isolation of brain ribonucleic acid

1979 ◽  
Vol 183 (1) ◽  
pp. 181-184 ◽  
Author(s):  
B B Kaplan ◽  
S L Bernstein ◽  
A E Gioio
Keyword(s):  
Rat Brain ◽  
Ribonucleic Acid ◽  
Guanidine Thiocyanate ◽  
Improved Method ◽  
Step Method ◽  
Rapid Isolation ◽  
Frozen Tissue ◽  
One Step ◽  
Protein Denaturant

An improved one-step method for the extraction of RNA from rat brain is described. Fresh or frozen tissue is disrupted in the powerful protein denaturant guanidine thiocyanate, and RNA isolated by ultracentrifugation through CsCl. The procedure is advantageous in that it is relatively simple, is rapid and does not expose the sample to enzyme treatments or repeated organic extractions.

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