Total RNA was isolated using the RNeasy Mini Kit (Qiagen) and DNaseI treated (Roche) according to manufacturer’s protocol. The High-Capacity cDNA reverse transcription kit (Applied Biosystems) was used to make cDNA, and PCR analysis was performed using the following primer sets: Vangl1 (5′-GGACTCAAGCCACAACGAGTTGTAT-3′) and (5′-ACTACGAGGCTGAAGTCCAAGC-3′), and Vangl2 (5′-ATGAGCGGGATGACAACTGG-3′ and 5′-ACCTTGAGCGTGAACTGAGG-3′). Quantitative real time PCR was performed in a Bio-Rad iCycler CFX-96 real-time PCR machine using SsoAdvanced Universal Probes Supermix (BioRad) and TaqMan gene-specific primer/probe sets (Applied Biosystems). Levels for VANGL1 (Hs01572998_m1), WNT5A (Hs00998537_m1), and NRDP1/RNF41 (Hs00195064_m1) were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 4352934E) levels for each sample.
Taqman gene specific primer probe sets
Manufactured by Thermo Fisher Scientific
TaqMan gene-specific primer/probe sets are research tools used in qPCR (quantitative Polymerase Chain Reaction) experiments. They consist of a pair of oligonucleotide primers and a fluorescently labeled TaqMan probe designed to detect and quantify specific DNA sequences.
Automatically generated - may contain errors
Lab products found in correlation
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (3 mentions)
Rneasy mini kit, by Qiagen (2 mentions)
Ssoadvanced universal probes supermix, by Bio-Rad (2 mentions)
Dnase 1, by Roche (2 mentions)
Icycler cfx 96, by Bio-Rad (2 mentions)
Cfx manager software, by Bio-Rad (1 mentions)
Ssoadvanced master mix, by Bio-Rad (1 mentions)
Cfx96 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Purelink rna mini kit, by Thermo Fisher Scientific (1 mentions)
3 protocols using taqman gene specific primer probe sets
1
Quantitative Analysis of VANGL Genes
2
Gene Expression Analysis via qPCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
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RNA was collected using a PureLink RNA MiniKit (Ambion) and converted to cDNA with the High-Capacity cDNA reverse transcription kit (Applied Biosystems). qPCR was conducted in a Bio-Rad CFX96 real-time PCR system using TaqMan gene-specific primer/probe sets (Applied Biosystems) and SsoAdvanced master mix (Bio-Rad). Analysis was conducted using Bio-Rad CFX Manager software, and message levels were normalized to GAPDH.
3
Quantitative Analysis of VANGL Genes
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