superscript ii™ reverse transcriptase reaction mix  (Thermo Fisher)

Article Title: Prostaglandin Analogs and Eupatilin as Treatments for Nephronophthisis
Article Snippet: .. Functional analyses were carried out using Metascape (http://metascape.org/). qRT-PCR Analysis Total RNA was reverse transcribed using SuperScript II Reverse Transcriptase (LifeTechnologies) according to the manufacturer’s protocol. .. Quantitative real-time polymerase chain reaction (PCR) (qRT-PCR) was performed with iTaq Universal SYBR Green Supermix (Bio-Rad) on the CFX-384 Real-Time PCR System (Biorad).

Article Title: Prostaglandin Analogs and Eupatilin as Treatments for Nephronophthisis
Article Snippet: .. Functional analyses were carried out using Metascape (http://metascape.org/). qRT-PCR Analysis Total RNA was reverse transcribed using SuperScript II Reverse Transcriptase (LifeTechnologies) according to the manufacturer’s protocol. .. Quantitative real-time polymerase chain reaction (PCR) (qRT-PCR) was performed with iTaq Universal SYBR Green Supermix (Bio-Rad) on the CFX-384 Real-Time PCR System (Biorad).

Article Title: Prostaglandin Analogs and Eupatilin as Treatments for Nephronophthisis
Article Snippet: .. Functional analyses were carried out using Metascape (http://metascape.org/). qRT-PCR Analysis Total RNA was reverse transcribed using SuperScript II Reverse Transcriptase (LifeTechnologies) according to the manufacturer’s protocol. .. Quantitative real-time polymerase chain reaction (PCR) (qRT-PCR) was performed with iTaq Universal SYBR Green Supermix (Bio-Rad) on the CFX-384 Real-Time PCR System (Biorad).

Article Title: Cucurbitacin IIa Alleviates Colitis via Promoting the Release of Host-Derived Extracellular Vesicles Encapsulating microRNA-30b-5p
Article Snippet: After evaluating RNA amount, purity, and fragment integrity, total RNA was first ligated with 3′ adapters using T4 RNA Ligase 2, followed by ligation with 5′ adapters using T4 RNA Ligase 1 (NEB, Phoenix, AZ, USA). .. Reverse transcription was performed with SuperScript II Reverse Transcriptase (THERMO, Shanghai, China) to produce cDNA, which was then amplified by PCR using High-Fidelity DNA Polymerase (NEB). .. The PCR products were purified and enriched through PAGE electrophoresis, followed by sequencing with Single-end 50 bp on the Illumina HiSeq 2500.

Article Title: Identification of a new COQ4 spliceogenic variant causing severe primary coenzyme Q deficiency.
Article Snippet: Background and aims: Primary Coenzyme Q (CoQ) deficiency caused by COQ4 defects is a clinically heterogeneous mitochondrial condition characterized by reduced levels of CoQ10 in tissues.. Next-generation sequencing has lately boosted the genetic diagnosis of an increasing number of patients.. Still, functional validation of new variants of uncertain significance is essential for an adequate diagnosis, proper clinical management, treatment, and genetic counseling.

Article Title: Myocarditis and neutrophil-mediated vascular leakage but not cytokine storm associated with fatal murine leptospirosis.
Article Snippet: .. One μg of RNA was subjected to reverse transcription using Superscript II reverse transcriptase (Invitrogen) according to the manufacturer’s recommendations. .. Generated cDNAs were either stored at −20 ◦C or used for qPCR on a StepOne Plus real-time PCR machine (Applied Biosystems) with primers and probes targeting murine hypoxanthine-guanine phosphoribosyltransferase (Hprt), used as a housekeeping gene, Il1b, Il6, Il10, Ifng, Nos2, Tnf, and Ccl5 (Table 2).

Article Title: Effective in vivo binding energy landscape illustrates kinetic stability of RBPJ-DNA binding
Article Snippet: To purify the total RNA, we used the RNeasy Mini Kit (Qiagen #74104), the QIAshredder (Qiagen #79654) and the DNase I (Qiagen #79254) accordingly to the manufacturer ́s instructions. .. For generation of cDNA, we used 1 μg of RNA and retro-transcribed it using random primers and SuperScript II reverse transcriptase (Invitrogen #18064-014). .. We assembled qPCR reactions with QuantiFast SYBR Green RT-PCR Kit (Qiagen #204156), gene-specific oligonucleotides (Supplementary Table ) and analyzed using the LightCycler480 TM system (Roche Diagnostics).

Article Title: Estrogen-Related Receptor is Required in Adult Drosophila Females for Germline Stem Cell Maintenance
Article Snippet: .. Cells were lysed in 250 μl lysis buffer from the RNAqueous-4PCR DNA-free RNA isolation for RT-PCR kit (Invitrogen), and RNA was extracted following the manufacturer’s instructions from three independent experiments. cDNA was synthesized from 500 ng of total RNA extracted from whole females or female adipocytes described above using Superscript II Reverse Transcriptase (Thermo Fisher Scientific) according to the manufacturer’s instructions. ..

Article Title: Cost-reduction strategy to culture patient derived bladder tumor organoids
Article Snippet: Total RNA was isolated from tumor organoids using TRIzol (ThermoFisher) on 10–14 days after culturing in both conditions (standard culture and cultured with alginate), and residual genomic DNA was digested with DNase I (Life Technologies). .. The cDNA was synthesized using Superscript II reverse transcriptase (Life Technologies) by using random primers. .. RT-quantitative PCRs (RT-qPCRs) were performed on a CFX Connect real-time PCR detection system thermocycler (Bio-Rad) using GoTaq qPCR master mix (Promega) (3 min at 95 °C, followed by 40 cycles of 95 °C for 10 s and 60 °C for 30 s).

Article Title: Cucurbitacin IIa Alleviates Colitis via Promoting the Release of Host-Derived Extracellular Vesicles Encapsulating microRNA-30b-5p
Article Snippet: After evaluating RNA amount, purity, and fragment integrity, total RNA was first ligated with 3′ adapters using T4 RNA Ligase 2, followed by ligation with 5′ adapters using T4 RNA Ligase 1 (NEB, Phoenix, AZ, USA). .. Reverse transcription was performed with SuperScript II Reverse Transcriptase (THERMO, Shanghai, China) to produce cDNA, which was then amplified by PCR using High-Fidelity DNA Polymerase (NEB). .. The PCR products were purified and enriched through PAGE electrophoresis, followed by sequencing with Single-end 50 bp on the Illumina HiSeq 2500.

Article Title: Cucurbitacin IIa Alleviates Colitis via Promoting the Release of Host-Derived Extracellular Vesicles Encapsulating microRNA-30b-5p
Article Snippet: After evaluating RNA amount, purity, and fragment integrity, total RNA was first ligated with 3′ adapters using T4 RNA Ligase 2, followed by ligation with 5′ adapters using T4 RNA Ligase 1 (NEB, Phoenix, AZ, USA). .. Reverse transcription was performed with SuperScript II Reverse Transcriptase (THERMO, Shanghai, China) to produce cDNA, which was then amplified by PCR using High-Fidelity DNA Polymerase (NEB). .. The PCR products were purified and enriched through PAGE electrophoresis, followed by sequencing with Single-end 50 bp on the Illumina HiSeq 2500.

Article Title: Ionizing radiation-induced disruption of Rela-Bclaf1-spliceosome regulatory axis in primary spermatocytes causing spermatogenesis dysfunction.
Article Snippet: The resulting fragments were reverse-transcribed to cDNA using SuperScriptTM II Reverse Transcriptase (Invitrogen, USA) and were converted into double-stranded DNA with E. coli (See figure on previous page.)

Article Title: Estrogen-Related Receptor is Required in Adult Drosophila Females for Germline Stem Cell Maintenance
Article Snippet: .. Cells were lysed in 250 μl lysis buffer from the RNAqueous-4PCR DNA-free RNA isolation for RT-PCR kit (Invitrogen), and RNA was extracted following the manufacturer’s instructions from three independent experiments. cDNA was synthesized from 500 ng of total RNA extracted from whole females or female adipocytes described above using Superscript II Reverse Transcriptase (Thermo Fisher Scientific) according to the manufacturer’s instructions. ..

Article Title: Estrogen-Related Receptor is Required in Adult Drosophila Females for Germline Stem Cell Maintenance
Article Snippet: .. Cells were lysed in 250 μl lysis buffer from the RNAqueous-4PCR DNA-free RNA isolation for RT-PCR kit (Invitrogen), and RNA was extracted following the manufacturer’s instructions from three independent experiments. cDNA was synthesized from 500 ng of total RNA extracted from whole females or female adipocytes described above using Superscript II Reverse Transcriptase (Thermo Fisher Scientific) according to the manufacturer’s instructions. ..

Article Title: Estrogen-Related Receptor is Required in Adult Drosophila Females for Germline Stem Cell Maintenance
Article Snippet: .. Cells were lysed in 250 μl lysis buffer from the RNAqueous-4PCR DNA-free RNA isolation for RT-PCR kit (Invitrogen), and RNA was extracted following the manufacturer’s instructions from three independent experiments. cDNA was synthesized from 500 ng of total RNA extracted from whole females or female adipocytes described above using Superscript II Reverse Transcriptase (Thermo Fisher Scientific) according to the manufacturer’s instructions. ..

Article Title: Estrogen-Related Receptor is Required in Adult Drosophila Females for Germline Stem Cell Maintenance
Article Snippet: .. Cells were lysed in 250 μl lysis buffer from the RNAqueous-4PCR DNA-free RNA isolation for RT-PCR kit (Invitrogen), and RNA was extracted following the manufacturer’s instructions from three independent experiments. cDNA was synthesized from 500 ng of total RNA extracted from whole females or female adipocytes described above using Superscript II Reverse Transcriptase (Thermo Fisher Scientific) according to the manufacturer’s instructions. ..

Article Title: Cost-reduction strategy to culture patient derived bladder tumor organoids
Article Snippet: Total RNA was isolated from tumor organoids using TRIzol (ThermoFisher) on 10–14 days after culturing in both conditions (standard culture and cultured with alginate), and residual genomic DNA was digested with DNase I (Life Technologies). .. The cDNA was synthesized using Superscript II reverse transcriptase (Life Technologies) by using random primers. .. RT-quantitative PCRs (RT-qPCRs) were performed on a CFX Connect real-time PCR detection system thermocycler (Bio-Rad) using GoTaq qPCR master mix (Promega) (3 min at 95 °C, followed by 40 cycles of 95 °C for 10 s and 60 °C for 30 s).