Total RNA of HT-1080 cells and tumor tissues was extracted by using an RNA Purified Total RNA Extraction Kit (Beyotime), and then reversely transcribed into cDNA with RT Master Mix (MedChemExpress, Monmouth Junction, NJ, USA), as we previously described (24 (link)). qPCR was performed using a SYBR Green qPCR kit (MedChemExpress). Specific primer sets were provided in Supplementary Table S1
Rt master mix
Manufactured by MedChemExpress
Sourced in United States
RT Master Mix is a ready-to-use solution containing all the necessary components for reverse transcription (RT) reactions. It provides an efficient and reliable way to synthesize cDNA from RNA templates.
Automatically generated - may contain errors
Lab products found in correlation
Sybr green qpcr master mix, by MedChemExpress (2 mentions)
Sybr green qpcr kit, by MedChemExpress (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Dna extraction kit, by Tiangen Biotech (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Sangon (1 mentions)
Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions)
Thiazolyl blue tetrazolium bromide, by Sangon (1 mentions)
Bacterial total rna extraction kit, by Tiangen Biotech (1 mentions)
4 protocols using rt master mix
1
Comprehensive RNA Extraction and qPCR Analysis
2
Quantitative PCR Analysis of RNA Expression
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Total RNAs were isolated from macrophages and corneal tissue using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and RNAs were reverse-transcribed into complementary DNAs (cDNAs) by RT-Master-Mix (MedChemExpress, Monmouth Junction, NJ, USA) following the manufacturer’s instructions. As a template for PCR, the cDNA was used with the SYBR-Green-qPCR Master Mix (MedChemExpress, Monmouth Junction, NJ, USA) on an ABI 7500 RT-PCR (Applied Biosystems, Foster City, CA, USA) thermocycler. After relative expressions were normalized to GAPDH, the data were analyzed using the 2−ΔΔCT method. Primers were designed by Sangon Biotech Company (Shanghai, China) and are shown in Table 1 .
3
Antimicrobial Activity Assay Protocol
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Luria–Bertani (LB) broth powder was purchased from Meilunbio (Dalian, China). 666-15 was purchased from Topscience (Shanghai, China). Antibiotics were purchased from MedChem Express (MCE, United States). Thiazolyl blue tetrazolium bromide (MTT) and phosphate-buffered saline (PBS) were purchased from Sangon Biotech (Shanghai, China). The bacterial total RNA extraction kit and DNA extraction kit were purchased from TIANGEN (Beijing, China). RT Master Mix and SYBR Green qPCR Master Mix were purchased from MedChem Express (MCE, United States). The lipopolysaccharide (LPS) detection kit was purchased from Cloud-Clone (Wuhan, China). Propidium iodide (PI) was purchased from Thermo Fisher Scientific, USA. 3,3-Dipropylthiadi-carbocyanine iodide [DiSC3(5)] was purchased from AAT Bioquest, USA. The LIVE/DEAD BacLight bacterial viability kit was purchased from Invitrogen, USA.
4
Gene Expression Analysis via qRT-PCR
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Cell/Tissue Total RNA Isolation Kit (RK02009, Biomarker,) was utilized to isolate the total RNAs, and cDNA synthesis was then operated using RT Master Mix (HY‐K0510A, MedChemExpress,). QRT‐PCR was utilized for detecting the mRNA expression levels of LIFR‐AS1, proliferating cell nuclear antigen (PCNA), E‐cadherin, N‐cadherin, and Snail using SYBR Green Fast qPCR Mix (RK02001, Biomarker,) in a D10 PCR gene amplification instrument (XuSensmart,). PCR conditions were as follows: 40 cycles of denaturation at 95°C for 20 s(s), annealing at 58°C for 20 s, and extension at 72°C for 20 s. The sequences of the primers are listed in Table 2 . Glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) was chosen as the internal control to normalize the gene expressions using the 2−ΔΔCT method.27
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