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Purelink micro to midi total rna purification system kit

Manufactured by Thermo Fisher Scientific
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The PureLink Micro-to-Midi Total RNA Purification System kit is a product designed for the isolation and purification of total RNA from a variety of sample types, including cells, tissues, and microorganisms. The kit utilizes spin column technology to efficiently capture and purify RNA, enabling users to obtain high-quality RNA for downstream applications.

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Lab products found in correlation

Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Abi prism 7900ht sequence detection system, by Thermo Fisher Scientific (1 mentions) Superscript double stranded cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) Superscript 3 platinum two step qrt pcr kit with sybr green, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

PureLink™ Micro-to-Midi Total RNA Purification System PureLink Micro-to-Midi total RNA purification kit Micro-to-Midi total RNA purification system Micro-to-Midi Total RNA Purification Kit PureLink RNA-micro-to-midi Kit PureLink Micro-to-Midi System Purelink micro to midi kit PureLink Total RNA Purification System PureLink micro RNA purification kit PureLink Total RNA purification kit

4 protocols using purelink micro to midi total rna purification system kit

1

SARS-CoV-2 Genes mRNA Expression in Transfected Cells

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To measure the mRNA co-expression of SARS-CoV-2 WH1 S, E, and M genes in transfected cells, the total RNAs were extracted from the stable transfected cells with pcDNA3.1-SARS-CoV-2 WH1 S-T2A-E-F2A-M using the PureLink Micro-to-Midi Total RNA Purification System kit (Invitrogen, Carlsbad, CA, USA) and synthesis cDNA with SuperScript™ III Reverse Transcriptase (Invitrogen, Carlsbad, CA, USA), and then evaluated using SYBR Green-based real-time PCR assay with SARS-CoV-2 WH1 S, E, and M-specific primer pairs. The S-specific primer pair had forward primer 5′-GCAATGAAGCCCAGCCAGATGTA-3′and reverse primer 5′-GTGGCTAAGAACCTGAATGAG-3′. The E-specific primers were forward primer 5′-ACAGGTACGTTAATAGTTAATAGCGT-3′ and reverse primer 5′-ATATTGCAGCAGTACGCACACA-3′. The M-specific primer pair contained forward primer 5′-TTTGTGCTTGCTGCTGTTTAC-3′ and reverse primer 5′-GAGTGGCACGTTGAGAAGAAT-3′. In addition, the glyceraldehyde 3-phosphate dehydrogenase (GAPDH)-specific primers pairs were forward primer 5′-TGCACCACCAACTGCTTAG-3′ and reverse primer 5′-GATGCAGGGATGATGTTC-3′. Finally, relative mRNA expression levels of SARS-CoV-2 WH1 S, E, and M genes in transfected cells were normalized by the GAPDH expression.
Chang Y.S., Chu L.W., Chen Z.Y., Wu J.S., Su W.C., Yang C.J., Ping Y.H, & Lin C.W. (2022). Development of Fluorescence-Tagged SARS-CoV-2 Virus-like Particles by a Tri-Cistronic Vector Expression System for Investigating the Cellular Entry of SARS-CoV-2. Viruses, 14(12), 2825.
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2

Evaluating SARS-CoV-2 PLpro and Fibrosis Genes

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To measure the expression of SARS PLpro, TGF-β1, pro-fibrotic and pro-protein convertase genes in transfected cells or mouse lung tissues, total RNAs were extracted from transfected A549 cells with empty vector pcDNA3.1 or pSARS-PLpro 2 days post transfection using PureLink Micro-to-Midi Total RNA Purification System kit (Invitrogen). Relative mRNA levels were analyzed using two-step real time RT-PCR with SYBR Green I, as described in our prior reports11 (link)12 (link). Primer pairs of SARS PLpro, TGF-β1, pro-fibrotic and pro-protein convertase genes were listed in Table 1. Quantification of specific PCR products was performed using the ABI Prism 7900HT Sequence Detection System (PE Applied Biosystems). Relative changes in mRNA level of indicated genes were normalized relative to GAPDH mRNA.
Li S.W., Wang C.Y., Jou Y.J., Yang T.C., Huang S.H., Wan L., Lin Y.J, & Lin C.W. (2016). SARS coronavirus papain-like protease induces Egr-1-dependent up-regulation of TGF-β1 via ROS/p38 MAPK/STAT3 pathway. Scientific Reports, 6, 25754.
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3

Extraction and Amplification of RNA Splice Variants

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Total RNA was isolated from all tissue samples using 2-mercaptoethanol and purified using the PureLink Micro-to-Midi Total RNA Purification System kit (Invitrogen).
Reverse transcription of RNA was performed using a Superscript Double-Stranded cDNA Synthesis Kit from Invitrogen with oligo primers. Forward and reverse primers used for initial PCR amplification of the splice variants are diagramed in figure 2 as follows: P1 = 5′-CTGCGTCTCCAAGATCCTCT-3′; P2 = 3′-AAAAGCCATCAGTTGGTTGG-5′; P3 = 5′-AGCAACTGGAAGAGCTGGAA-3′; P4 = 3′-TGACAGGGTCCATGCTGTAG-5′.
Adams J.S., Sudweeks S.N, & Stark M.R. (2014). Pax3 isoforms in sensory neurogenesis: expression and function in the ophthalmic trigeminal placode. Developmental dynamics : an official publication of the American Association of Anatomists, 243(10), 1249-1261.
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4

Quantitative Analysis of Host Chemokines and Viral Proteins

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The total RNAs were purified from transfected cells, stable cell lines and mouse lung tissues using the PureLink Micro-to-Midi Total RNA Purification System kit (Invitrogen, Carlsbad, CA, USA). Relative mRNA levels of host chemokines and viral SUD were analyzed using a SuperScript™ III Platinum® Two-Step qRT-PCR Kit with SYBR® Green (Invitrogen, Carlsbad, CA, USA). After reverse transcription by SuperScript III RT, the real-time PCR was performed with the cDNAs, specific primer pairs, and Platinum® SYBR® Green qPCR SuperMix according to the amplification protocol consisting of 1 cycle at 50 °C for 2 min, 1 cycle at 95 °C for 10 min, 45 cycles at 95 °C for 15 sec, and 60 °C for 1 min. Primer pairs for the detection mRNA levels of human and mouse chemokines (CXCL8, CXCL10, CXCR3, CCL-2/MCP-1, CCL3/MIP-1α, and CCL5/RANTES) as well as SARS-CoV SUD, SUD-NM, and SUD-MC are listed in Table 2. Relative mRNA levels of indicated genes were standardized by human β-actin and mouse GAPDH and then calculated by the comparative CT method (ΔΔCT method).
Chang Y.S., Ko B.H., Ju J.C., Chang H.H., Huang S.H, & Lin C.W. (2020). SARS Unique Domain (SUD) of Severe Acute Respiratory Syndrome Coronavirus Induces NLRP3 Inflammasome-Dependent CXCL10-Mediated Pulmonary Inflammation. International Journal of Molecular Sciences, 21(9), 3179.
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