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Purelink micro to midi system

Manufactured by Thermo Fisher Scientific
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The PureLink Micro-to-Midi System is a versatile DNA/RNA purification tool designed for small- to medium-scale nucleic acid extraction. It utilizes a silica-based membrane technology to efficiently purify DNA or RNA from a variety of sample types.

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Lab products found in correlation

Primescript rt reagent kit, by Takara Bio (4 mentions) Sequence detection system software, by Thermo Fisher Scientific (3 mentions) 7500 real time pcr system, by Thermo Fisher Scientific (3 mentions) Sybr premix ex taqtm, by Takara Bio (2 mentions) Sybr premix ex taq, by Takara Bio (2 mentions) 7500 real time pcr system, by Takara Bio (1 mentions) Purelink ffpe rna isolation kit, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Collagenase 2, by Worthington (1 mentions) Rt2 qpcr primer assay, by Qiagen (1 mentions)

Close spelling variants of this product

PureLink Micro-to-Midi Total RNA Purification System kit PureLink™ Micro-to-Midi Total RNA Purification System

5 protocols using purelink micro to midi system

1

Real-Time PCR Quantification of Gene Expression

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Total RNA was extracted using the PureLinkTM Micro-to-Midi System (Invitrogen) according to the manufacturer’s instructions, and reverse transcription was used to generate cDNAs using the PrimeScriptTM RT Reagent kit (TaKaRa). Real time PCR was performed using SYBR Premix Ex TaqTM (TaKaRa) and the 7500 Real-Time PCR System (Applied Biosystems). The reaction parameters were 95 °C for 30 s followed by 40 two-step cycles of 95 °C for 5 s and 60 °C for 34 s. All the primer pairs used to PCR amplification were shown in Supplementary Table S5. Ct values were calculated using Sequence Detection System software (Applied Biosystems), and the amount of target sequence normalized to the reference sequence was calculated as 2−ΔΔCt.
Li J., Gao Z., Wang X., Liu H., Zhang Y, & Liu Z. (2016). Identification and functional analysis of long intergenic noncoding RNA genes in porcine pre-implantation embryonic development. Scientific Reports, 6, 38333.
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2

Quantitative Real-Time PCR of Oocyte RNA

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Total RNA was extracted from at least 100 oocytes using the PureLinkTM Micro-to-Midi system (Invitrogen) according to the manufacturer’s instructions, and reverse transcription was used to generate cDNAs using the PrimeScriptTM RT Reagent Kit (TaKaRa). Real-time PCR was performed using SYBR Premix Ex TaqTM (TaKaRa) and the 7500 Real-Time PCR System (Applied Biosystems). The reaction parameters were 95 °C for 30 sec, followed by 40 two-step cycles of 95 °C for 5 sec and 60 °C for 34 sec. Primers for each gene were listed (Additional file 1: Table S1). 18S rRNA was used as a reference gene. Ct values were calculated using Sequence Detection System software (Applied Biosystems), and the amount of target sequence normalized to the reference sequence was calculated as 2–△△Ct.
Zhao Q., Guo Z., Piao S., Wang C, & An T. (2015). Discovery of porcine maternal factors related to nuclear reprogramming and early embryo development by proteomic analysis. Proteome Science, 13, 18.
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3

Total RNA Extraction and qPCR Analysis

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Total RNA was extracted using the PureLink Micro-to-Midi System (Invitrogen) according to the manufacturer’s instructions, and reverse transcription was used to generate cDNAs using the PrimeScript RT Reagent kit (TaKaRa). qPCR was performed using SYBR Premix Ex Taq (TaKaRa) and the 7,500 Real-Time PCR System. The reaction parameters were 95°C for 30 s, followed by 40 two-step cycles of 95°C for 5 s and 60°C for 34 s. Ct values were calculated using Sequence Detection System software, and the amount of target sequence normalized to the reference sequence was calculated as 2−ΔΔCt. All primers and probes were designed using Primer Premier 5.
Yang X., Ji J., Cui H., Zhao Q., Ding C, & Xu C. (2022). Functional evaluation of LTR-derived lncRNAs in porcine oocytes and zygotes with RNA-seq and small RNA-seq. Frontiers in Genetics, 13, 1023041.
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4

Quantitative Transcriptome Analysis of Immune Cells in Myocarditis

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Mouse hearts were perfused and digested with collagenase II (10,000 U/mL, Worthington, CLS-2) to release immune cells using a gentleMACs Dissociator. Immune cells were isolated using MACs magnetic microbeads for CD11b or CD45 (Miltenyi Biotec, Auburn, CA). Trizol Reagent and the PureLink Micro-to-Midi system (Invitrogen, Carlsbad, CA) were used for purification of RNA from total hearts or immune cells. Human RNA was isolated from heart biopsies of formalin-fixed, paraffin-embedded (FFPE) blocks from patients with histology-verified myocarditis. Human RNA was processed using a PureLink FFPE RNA Isolation Kit (Invitrogen, Carlsbad, CA). Quantitative RT-PCR (qRT-PCR) of gene expression was measured using Assay-on-Demand probe sets (Applied Biosystems, Foster City, CA) or RT2qPCR Primer Assay (Qiagen, Venlo, Netherlands) and reactions analyzed using the ABI 7000 Taqman system, as previously [13 (link), 21 (link)]. Hypoxanthine phosphoribosyltransferase (Hprt) was used for normalization.
Fairweather D., Coronado M.J., Garton A.E., Dziedzic J.L., Bucek A., Cooper LT J.r., Brandt J.E., Alikhan F.S., Wang H., Endres C.J., Choi J., Pomper M.G, & Guilarte T.R. (2014). Sex differences in translocator protein 18 kDa (TSPO) in the heart: implications for imaging myocardial inflammation. Journal of cardiovascular translational research, 7(2), 192-202.
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5

Quantification of Gene Expression in Oocytes

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Total RNA was extracted using the PureLink Micro-to-Midi System (Invitrogen) according to the manufacturer's instructions and reverse transcription was used to generate cDNAs using the PrimeScript RT Reagent kit (TaKaRa). Real-time PCR was performed using SYBR Premix Ex Taq (TaKaRa) and the 7500 Real-Time PCR System (Applied Biosystems). The reaction parameters were 958C for 30 s followed by 40 two-step cycles of 958C for 5 s and 608C for 34 s. All the primer pairs used for PCR amplification are shown in Table S1, available as Supplementary Material to this paper. Ct values were calculated using Sequence Detection System software (Applied Biosystems) and the amount of target sequence normalised to the reference sequence was calculated by the DD CT method. The 18SrRNA was used as control. The oocytes without injection were used as a reference sample. Each pool contained 50 oocytes. All data are based on biological triplicates.
Liu W., Zhao Q., Piao S., Wang C., Kong Q, & An T. (2017). Endo-siRNA deficiency results in oocyte maturation failure and apoptosis in porcine oocytes. Reproduction, fertility, and development, 29(11).
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