Poly gel rna extraction kit

Manufactured by Omega Bio-Tek

Sourced in China

The Poly-Gel RNA Extraction Kit is a laboratory tool designed for the extraction and purification of RNA from a variety of sample types. The kit utilizes a polymer-based gel matrix to efficiently capture and isolate RNA molecules, enabling their subsequent analysis and downstream applications.

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Lab products found in correlation

T7 rna polymerase, by Promega (4 mentions) Rq1 rnase free dnase 1, by Promega (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Quantity one, by Bio-Rad (1 mentions)

Spelling variants of this product

Gel Extraction Kit (124 citations) RNA extraction kit (50 citations)

4 protocols using poly gel rna extraction kit

In vitro Transcription and Purification of FHV RNA1

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The DNA fragment of the 3′-end of the (+)RNA1, (–)RNA1 was amplified from the cDNA of the FHV RNA1 genome plasmids FHV1(1, 0). Following the manufacturer’s instructions (Promega, Madison, WI), each pair of the primers was designed to include one primer, which contains a portion of 5′-end of expected RNA product plus the T7 promoter region (TAATACGACTCACTATAGg), and the other primer whose sequence is reverse-complementary to the 3′-end of the expected RNA product. RNA was transcribed in vitro from PCR products using T7 RNA polymerase (Promega, Madison, WI) for 4 h. DNA templates were removed by RQ1 RNase-free DNase I (Promega) at 37°C for 1 h. After that, TRIzol reagent (Invitrogen) was used to extract the RNAs according to the manufacturer’s protocol. The blocking of RNA template was conducted as previously described [41] (link).
To get rid of T7 RNA polymerase and NTP, all the RNA templates were electrophoresed in a 6% polyacrylamide–7 M urea gel and further purified by Poly-Gel RNA Extraction Kit (Omega bio-tek, Norcross, GA) according to the manufacturer’s instruction. Purified RNAs were quantified with Bio-Rad Quantity One using known amounts of RNA as controls after Northern blots.

Wu W., Wang Z., Xia H., Liu Y., Qiu Y., Liu Y., Hu Y, & Zhou X. (2014). Flock House Virus RNA Polymerase Initiates RNA Synthesis De Novo and Possesses a Terminal Nucleotidyl Transferase Activity. PLoS ONE, 9(1), e86876.

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Preparation of RNA and DNA Hybrid Substrates

Cited 1 time

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In brief, of the two strands, one was labeled at the 5′ end with hexachloro-fluorescein (HEX), and the other strand was unlabeled. HEX-labeled oligonucleotide strands were purchased from TaKaRa (Dalian, China). Unlabeled DNA strands were synthesized by Invitrogen, and unlabeled RNA strands were in vitro transcribed using T7 RNA polymerase (Promega, Madison, WI). The in vitro transcribed RNA strands were purified by Poly-Gel RNA Extraction Kit (Omega bio-tek, Guangzhou, China) according to the manufacturer's instruction. The two strands were mixed in a proper ratio, and annealed through heating and gradually cooling as previously described [30 (link),40 (link)]. The standard RNA helix substrate with both 5′ and 3′ protrusions was annealed with RNA1 and RNA2, the R*/D substrate was annealed with RNA1 and DNA1, the D*/D substrate was annealed with DNA2 and DNA3, the D*/R substrate was annealed with DNA2 and RNA3, the 3′-protruded RNA helix was annealed with RNA1 and RNA4, the 5′-protruded RNA helix was annealed with RNA1 and RNA5, the blunt-ended substrate was annealed with RNA1 and RNA6, and the 49 matched bps substrate was annealed with RNA7 and RNA8. All oligonucleotides used in this study are listed in S2 Table.

Xia H., Wang P., Wang G.C., Yang J., Sun X., Wu W., Qiu Y., Shu T., Zhao X., Yin L., Qin C.F., Hu Y, & Zhou X. (2015). Human Enterovirus Nonstructural Protein 2CATPase Functions as Both an RNA Helicase and ATP-Independent RNA Chaperone. PLoS Pathogens, 11(7), e1005067.

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RNA Helix Substrate Preparation

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The RNA helix substrates consist of two complementary nucleic acid strands, one of which was labeled at 5′-end with Hexachloro-Fluorescein hosphoramidite (HEX) and the other was unlabeled. The labeled strands were purchased from TaKaRa (Dalian, China). All the unlabeled RNA strands were transcribed in vitro using T7 RNA polymerase (Promega, Madison, WI). The posttranscriptional RNAs were purified by using Poly-Gel RNA extraction kit (Omega Bio-Tek, Guangzhou, China) according to the manufacturer′s instructions. The oligonucleotide helices were generated by annealing the labeled strand and unlabeled strand, at which mixed a 1:1 ratio in a 10-μL reaction mixture containing 25 mmol/L HEPES–KOH (pH 8.0) and 25 mmol/L NaCl. The mixture was heated to 95 °C for 5 min and was then cooled gradually to 25 °C to produce helical duplexes. Two RNA helix substrate with both 5′- and 3′-protrusions was annealed with RNA1 and RNA2 (24-nt non-labeled) or RNA3 (28-nt non-labeled). The 5′-protruded RNA helix was annealed with RNA1 and RNA4. The 3′-protruded RNA helix was annealed with RNA1 and RNA5. The blunt RNA helix was annealed with RNA1 and RNA6. All the oligonucleotides are listed in Supplementary Table S2.

Shu T., Huang M., Wu D., Ren Y., Zhang X., Han Y., Mu J., Wang R., Qiu Y., Zhang D.Y, & Zhou X. (2020). SARS-Coronavirus-2 Nsp13 Possesses NTPase and RNA Helicase Activities That Can Be Inhibited by Bismuth Salts. Virologica Sinica, 35(3), 321-329.

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RNA Helix Substrate Preparation

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The RNA helix substrates were composed of two complementary nucleic acid strands: one with a short hexachloro fluorescein (HEX) label at the 5′ end, and the other with no label. The labeled RNA strands were synthesized by TaKaRa (Dalian, China), while the unlabeled strands were produced by in vitro transcription using T7 RNA polymerase (Promega, Madison, WI, USA). The posttranscriptional RNA purification was performed with Poly-Gel RNA extraction kit (Omega Bio-Tek, Guangzhou, China) according to the manufacturer's protocol. The labeled RNA strands and the unlabeled RNA strands were mixed in the ratio of 1:1, and were added to a 10-μL reaction mixture containing 25 mmol/L HEPES-KOH (pH 8.0) and 25 mmol/L NaCl. Subsequently, the reaction mixture underwent an annealing process, which was specifically to heat the reaction mixture to 95 °C for 5 min, followed by a gradual cooling to 25 °C to produce helical duplexes. Two RNA helix substrates with both 5′- and 3′-protrusions were annealed with RNA1 and RNA2. The 3′-protruded RNA helix and 5′-protruded RNA helix were produced by annealing RNA1 with RNA3 and RNA4, respectively. Supplementary Table S2 shows all the oligonucleotides that have been used.

Chen Z., Xiong X., Li Y., Huang M., Ren Y., Wu D., Qiu Y., Chen M., Shu T, & Zhou X. (2022). The nonstructural protein 2C of Coxsackie B virus has RNA helicase and chaperoning activities. Virologica Sinica, 37(5), 656-663.

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