Rnazol rt reagent

Manufactured by GeneCopoeia

Sourced in United States

RNAzol RT is a reagent used for the isolation and purification of total RNA from various biological samples. It is a guanidinium thiocyanate-phenol-based solution that effectively lyses cells and denatures protein complexes to release RNA. The isolated RNA can then be used for various downstream applications, such as gene expression analysis, RT-PCR, and Northern blotting.

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Lab products found in correlation

Nanodrop nd 1000 uv vis spectrophotometer, by Thermo Fisher Scientific (1 mentions) Eco thermocycler, by Illumina (1 mentions) Surescript first strand cdna synthesis kit, by GeneCopoeia (1 mentions) Blazetaq sybr green qpcr mix 2, by GeneCopoeia (1 mentions)

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RNAzol® RT RNA Isolation Reagent (7 citations) RNAzol (6 citations) RNAzol reagent (3 citations)

4 protocols using rnazol rt reagent

Quantitative gene expression analysis in chickens

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The procedure was conducted as described in Dishon et al. (2017) (link). Briefly, frozen samples were homogenized, and total RNA was extracted using RNAzol RT reagent (Genecopoeia,Rockville, MD). Concentration was measured by NanoDrop ND-1000 UV-Vis spectrophotometer (NanoDrop Technologies, Wilmington, DE). A reverse transcription reaction was performed to produce a total 20 µL cDNA. Real-time PCR was conducted using β-actin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the standards. The sequences of all gene-specific primers used in the PCR are shown in Table 2. Gene-expression results are presented in arbitrary units (AU).Table 2

Primers used in real-time PCR.

Table 2

Gene	Primers (5′→3′)	Product length	GenBank accession no.
GAPDH	R:CCTGCATCTGCCCATTTF:GGCACGCCATCACTATC	61	NM 204,305.1
Actin	R:AAAGCCATGCCAATCTCGTCF:CCGCAAATGCTTCTAAACCG	101	NM 205,518.1
Aromatase	R:AGCTGTTAGGCAATACTGTGGAF:CCAGTTGCCACAGTGCCTAT	114
GnRH-1	R:GATCAGGCTTGCCATGGTTTF:TGGTCTTATGGCCTGCAACC	174	NM_001080877.1
LH	R:CCCCATAAGTGCACGCCGF:GAATGCCCCCAATGTATGGCT	118	S_70834
FSH	R:ATTCAGGATGGTCACCGCAGF:CTGCTTCACAAGGGATCCAGTA	122	NM_204257.1
LH receptor	R:GGTAGGTCAGAACGGCTTCCF:GGCCAGACGTCCTGGATATT	159	NM_204936.1
FSH receptor	R:GGACAAATCTCAGTTCTGTGGCF:GGACAAATCTCAGTTCTGTGGC	154	NM_205079.1
Serotonin transporter	R:CGCTTGCTCCAGACGTGTF: CCGAAGCCATTGCGAACA		NM_213572.1

Bartman J., Zaguri S., Avital-Cohen N., Dishon L., Druyan S., Gumułka M, & Rozenboim I. (2021). Targeted differential illumination improves reproductive traits of broiler breeder males. Poultry Science, 100(6), 101109.

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Quantification of Gene Expression in Frozen Tissue Samples

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The frozen tissue samples were homogenized by HG-300 homogenizer, and total RNA was extracted using RNAzol RT reagent, according to the manufacturer's protocol (GeneCopoeia, Rockville, MD). The RNA extraction protocol, as well as the cNDA production and real time PCR protocols was as described in our previous papers (Dishon et al., 2017 (link); Dishon et al., 2021). A list of primers is found in Table 1.Table 1

Primers used in real-time PCR reactions.

Table 1:

Gene	Primers	Product length	GenBank accession no.
GAPDH	F: GGCACGCCATCACTATC	61 bp	NM_204305.1
GAPDH	R: CCTGCATCTGCCCATTT	61 bp	NM_204305.1
β-Actin	F: CCGCAAATGCTTCTAAACCG	101 bp	NM_205518.1
β-Actin	R: AAAGCCATGCCAATCTCGTC	101 bp	NM_205518.1
GHRH	F: GGCAAACGGCTCAGAAACAG	140 bp	NM_001040464.1
GHRH	R: AGCATGGCTCCCAAGAAGTC	140 bp	NM_001040464.1
GHR	F: GCGTGTTCAGGAGCAAAGCT	121 bp	NM_001001293.1
GHR	R: TGGGACAGGCATTTCCATACTT	121 bp	NM_001001293.1
IGF	F: GCTTTTGTGATTTCTTGAAGGTGAA	195 bp	NM_001004384.2
IGF	R: CATACCCTGTAGGCTTACTGAAGTA	195 bp	NM_001004384.2

Dishon L., Avital-Cohen N., Zaguri S., Bartman J., Heiblum R., Druyan S., Porter T.E., Gumułka M, & Rozenboim I. (2021). The effect of selected in ovo green light photostimulation periods on post-hatch broiler growth and somatotropic axis activity. Poultry Science, 100(8), 101229.

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MicroRNA Expression Analysis in Cells and Tissues

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MiRNA was isolated from cultured cells or ventricular tissue using RNAzolRT reagent (GeneCopoeia, Rockville, MD, USA) and the samples were reverse transcribed using the All-in-One RT-qPCR kit (GeneCopoeia, Cat#: QP016) according to the supplier’s instructions. The cDNA samples were further processed through quantitative RT-qPCR using miRNA specific primer sets for 1a, 133, 27b, or 208a from GeneCopoeia. MiRNA expression for RT-qPCR analysis was normalized to a reference primer (U6) using the ΔΔC_T method. A thermal profile was utilized as follows: 50 °C for 2 min, 95 °C for 10 min, 95 °C for 10 sec, 63 °C for 20 sec, 72 °C for 10 sec; 40 cycles; then 95 °C, 60 °C and 95 °C for 15 sec each, using an ECO thermocycler (Illumina, San Diego, CA).

Govindapillai A., Hotchkiss A., Baguma-Nibasheka M., Rose R.A., Miquerol L., Smithies O., Maeda N, & Pasumarthi K.B. (2018). Characterizing the role of atrial natriuretic peptide signaling in the development of embryonic ventricular conduction system. Scientific Reports, 8, 6939.

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Gene Expression Analysis by qRT-PCR

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Total RNA was obtained using RNAzol®RT reagent (GeneCopoeia, USA) and then reversely transcribed into cDNA using the SureScript™ First‐Strand cDNA Synthesis Kit (GeneCopoeia). Quantitative real‐time PCR (qRT‐PCR) was carried out using the BlazeTaq™ SYBR® Green qPCR Mix 2.0 (GeneCopoeia). The relative gene levels were normalized to glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) and calculated by the 2^−ΔΔCt method. The primers used are listed in Table1.

Chen Z., He C., Gao Z., Li Y., He Q., Wang Y, & Cai C. (2023). Polypyrimidine tract binding protein 1 exacerbates cardiac fibrosis by regulating fatty acid‐binding protein 5. ESC Heart Failure, 10(3), 1677-1688.

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