Total RNA was isolated using Plant Total RNA kit (Invitrogen, USA). RNA quality and quantity were determined using a Nanodrop 2,000 spectrophotometer (Thermo Scientific, USA). Only RNA samples with A260/A280 between 1.8 and 2.1 and A260/A230 between 2.0 and 2.2 were used. Total RNA integrity was checked via 1.2% agarose gel electrophoresis under denaturing conditions. RNA samples were treated with RQ1 RNase-Free DNase (Promega, USA). cDNA was synthesized from 2 μg of total RNA using the SuperScriptTM RT kit (Thermo Scientific, USA) in a 20 μL- reaction volume according to the manufacturer's recommendations. cDNA was stored at −20°C until further use.
Plant total rna kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Plant Total RNA kit is a laboratory product designed to efficiently extract and purify total RNA from a variety of plant tissues. The kit utilizes a proven method to isolate high-quality RNA that can be used in downstream applications such as RT-qPCR, RNA sequencing, and other molecular biology techniques.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Superscript rt kit, by Thermo Fisher Scientific (1 mentions)
Rq1 rnase free dnase, by Promega (1 mentions)
Power cdna synthesis kit, by iNtRON Biotechnology (1 mentions)
Cfx96 real time system, by Bio-Rad (1 mentions)
Nanophotometer, by Implen (1 mentions)
2 protocols using plant total rna kit
1
Total RNA Isolation and cDNA Synthesis
2
Salinity Stress Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The Plant Total RNA kit (Invitrogen, USA) was used for the extraction of total RNA from leaf samples, according to the manufacturer's instructions. The nanophotometer (Implen Inc., Westlake) was used to ensure the quality and quantity of RNA while the ethidium-bromide stain analyses using agarose gel electrophoresis was used to check the purity of total RNA. The Power cDNA synthesis kit (Intron Biotechnology Inc., USA) was used to reverse transcribe DNA free total RNA into cDNA.
To evaluate the expression pattern of salinity stress candidate genes, antioxidant-related genes, osmolytes-biosynthesis-related, photosystemrelated and ABA-related genes used in the study (Table 1), the CFX 96 Real-Time system (Bio-Rad, Richmond, CA, USA) with SYBR-green fluorescence was used and analyses of the results were done by using the ΔΔCT method. The conditions for the thermal cycle was 95°C for 5min and 40 cycles of 95°C for 15s, 55°C for 15s and 72°C for 30s. The experiment was triplicated, using the Actin as an internal control for standardizing the relative transcript levels.
To evaluate the expression pattern of salinity stress candidate genes, antioxidant-related genes, osmolytes-biosynthesis-related, photosystemrelated and ABA-related genes used in the study (Table 1), the CFX 96 Real-Time system (Bio-Rad, Richmond, CA, USA) with SYBR-green fluorescence was used and analyses of the results were done by using the ΔΔCT method. The conditions for the thermal cycle was 95°C for 5min and 40 cycles of 95°C for 15s, 55°C for 15s and 72°C for 30s. The experiment was triplicated, using the Actin as an internal control for standardizing the relative transcript levels.
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