A total of 1.5 × 105 or 2 × 105 HFt cells seeded in 12-well plates were incubated at 37°C in 5% CO2 overnight or at least 4 h before further manipulation. Total RNA was isolated using the RNAeasy Plus kit (Qiagen; 74134) or TRIzol reagent (Invitrogen; 15596) according to the manufacturer's instructions. RNA was reverse transcribed using the TaqMan reverse transcription reagent kit (Life Technologies; N8080234) with oligo(dT) primers. Samples were analyzed in triplicate using TaqMan Fast Universal PCR master mix (Life Technologies; 4352042) with the following TaqMan gene-specific primer (6-carboxyfluorescein [FAM]/MGB)-probe mixes (Life Technologies): PML (assay ID Hs00231241_m1), PIAS4 (assay ID Hs00249203_m1), 18S (GenBank accession number X03205.1 ; Thermo Fisher Scientific; 4319413E), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH; GenBank accession number NM_002046.5 ; Thermo Fisher Scientific; 4333764F).
Taqman gene specific primer 6 carboxyfluorescein fam mgb probe mixes
Manufactured by Thermo Fisher Scientific
The TaqMan gene-specific primer (6-carboxyfluorescein [FAM]/MGB)-probe mixes are pre-designed and optimized oligonucleotides used for the detection and quantification of specific gene targets in real-time PCR and reverse transcription-PCR (RT-PCR) applications. The mixes include a forward primer, a reverse primer, and a fluorogenic probe labeled with FAM and a minor groove binder (MGB) that provides enhanced specificity and sensitivity.
Automatically generated - may contain errors
Lab products found in correlation
Rnaeasy plus kit, by Qiagen (2 mentions)
Taqman reverse transcription reagent kit, by Thermo Fisher Scientific (2 mentions)
Taqman fast universal pcr master mix, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Gapdh, by Thermo Fisher Scientific (2 mentions)
2 protocols using taqman gene specific primer 6 carboxyfluorescein fam mgb probe mixes
1
Quantifying mRNA Expression in HFt Cells
2
Quantitative Analysis of Gene Expression
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A total of 1.5 × 105 or 2.0 × 105 HFt cells seeded in 12-well plates were incubated at 37°C in 5% CO2 overnight, or for at least 4 h before further manipulation. Cells were washed with PBS and total RNA was isolated using the RNAeasy Plus kit (Qiagen; 74137) or TRIzol reagent (Invitrogen; 15596) by following the manufacturer's instructions. cDNA was synthesized using the TaqMan reverse transcription reagent kit (Life Technologies; N8080234) with oligo(dT) primers. Samples were analyzed in triplicate using TaqMan Fast Universal PCR master mix (Life Technologies; 4352042) with TaqMan gene-specific primer (6-carboxyfluorescein [FAM]/MGB)-probe mixes (Life Technologies) for the following: PML (Hs00231241_m1), PIAS1 (Hs00184008_m1), 18S (GenBank accession number X03205.1 ), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Life Technologies product code 4333764F).
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