qRT-PCR Analysis of HaCaT Keratinocyte mRNA

The mRNA levels of HaCaT keratinocytes were analyzed by qRT-PCR using the LightCycler 96 Instrument (Roche Molecular Systems Inc., Basel, Switzerland) as our previous report [10 (link),49 (link),50 (link)]. Total cellular RNA in HaCaT cells was extracted by using Easy Blue^® kits (Intron Biotechnology, Seoul, Korea), synthesis to cDNA using 0.5 mg/mL random oligonucleotide primers (Promega, Madison, WI, USA) and TOPscript^TM RTDryMIX (Enzynomics, Daejeon, Korea). PCR amplification was analyzed by using the incorporation of SYBR green (TaKaRa, Shiga, Japan) and specific primers. Primer sequences are listed in Table S1.

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Choi S.I., Han H.S., Kim J.M., Park G., Jang Y.P., Shin Y.K., Ahn H.S., Lee S.H, & Lee K.T. (2021). Eisenia bicyclis Extract Repairs UVB-Induced Skin Photoaging In Vitro and In Vivo: Photoprotective Effects. Marine Drugs, 19(12), 693.

Publication 2021

LightCycler 96 Instrument Easy Blue® kits random oligonucleotide primers TOPscriptTM RTDryMIX SYBR green

Cdna Cellular Hacat cells Intron Keratinocytes Mrna Primer Promega Sybr green Synthesis

Corresponding Organization : Kyung Hee University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

MRNA levels of HaCaT keratinocytes

control variables

LightCycler 96 Instrument (Roche Molecular Systems Inc., Basel, Switzerland) used for qRT-PCR analysis
Total cellular RNA in HaCaT cells extracted using Easy Blue® kits (Intron Biotechnology, Seoul, Korea)
CDNA synthesis using 0.5 mg/mL random oligonucleotide primers (Promega, Madison, WI, USA) and TOPscript™ RTDryMIX (Enzynomics, Daejeon, Korea)
PCR amplification analyzed using the incorporation of SYBR green (TaKaRa, Shiga, Japan) and specific primers

positive controls

None explicitly mentioned

negative controls

None explicitly mentioned

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