Total RNA from the gastrocnemius muscle tissue was isolated using PureLink RNA lysis buffer (Invitrogen, Carlsbad, CA, USA). Contaminated genomic DNA was digested with DNase I treatment (Invitrogen) at room temperature for 15 min. Complementary DNA was prepared from the total RNA using reverse transcriptase, according to the manufacturer's instructions (High Capacity cDNA Reverse Transcription Kit; Applied Biosystems, Foster City, CA, USA). Quantitative mRNA expression was assessed by real-time PCR using the Power SYBR Green PCR Master Mix (Applied Biosystems) and target gene-specific primers. All the primers are listed in the supplemental file (Table S1 ). Samples were analyzed using the 7500 Real-Time PCR system, and the data were analyzed using 7500 Software (Applied Biosystems). Relative expression levels of the WT + V target mRNA group were compared with those of the controls.
Purelink rna lysis buffer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PureLink RNA lysis buffer is a reagent used in the RNA extraction and purification process. It is designed to efficiently lyse cells and release their RNA content, facilitating the subsequent steps of RNA isolation.
Automatically generated - may contain errors
Lab products found in correlation
Dnase 1 treatment, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
7500 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Power sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions)
Acculan 3ti, by B. Braun (1 mentions)
2 protocols using purelink rna lysis buffer
1
Quantitative mRNA Expression Analysis
2
Isolation and FACS Sorting of Dermal Cell Populations
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were isolated from a 10 Â 10-cm human skin piece from whole skin and/or dermatomed (Acculan 3Ti; Aesculap, Center Valley, PA) papillary (300 mm) and upper (300e1,000 mm) and lower (>1,000 mm) reticular dermis after subcutaneous fat removal and separation of epidermis using dispase II with a digestion cocktail, as previously described (Lichtenberger et al., 2016) . Single-cell suspensions were stained for FACS according to standard procedures. FACS-sorted cells were collected in PureLink RNA-lysis buffer (Invitrogen, Waltham, MA) or in medium for tissue culture.
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