Total RNA was extracted from cells using the Blood/Cultured Cell Total RNA Mini Kit (FAVORGEN, Ping Tung, Taiwan), followed by reverse transcription using ReverTra Ace qPCR RT Master Mix (Toyobo, Osaka, Japan) according to the manufacturer’s instructions. cDNA was amplified for 40 cycles in a Light Cycler 480 (Roche) using LightCycler 480 SYBR Green I Master reagent (Roche). Expression of survivin was normalized to that of GAPDH mRNA (internal standard) by the ΔΔCt method. The primer pairs were as follows (final concentration 0.5 μM): human survivin, 5′-GGACCACCGCATCTCTACAT-3′ and 5′-GC ACTTTCTTCGCAGTTTCC-3′; human GAPDH, 5′-GAAA GGTGAAGGTCGGAGTC-3′ and 5′-GAAGATGGTGATG GGATTTC-3′. Three independent experiments were performed.
Blood cultured cell total rna mini kit
Manufactured by Favorgen Biotech
The Blood/Cultured Cell Total RNA Mini Kit is a laboratory equipment designed to efficiently extract and purify total RNA from whole blood samples or cultured cells. The kit utilizes a silica-based membrane technology to capture and wash the RNA, ensuring high-quality RNA is obtained for downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Lightcycler 480, by Roche (3 mentions)
Revertra ace qpcr rt master mix, by Toyobo (2 mentions)
Lightcycler 480 sybr green 1 master reagent, by Roche (1 mentions)
Revertra ace, by Toyobo (1 mentions)
Primescript real time pcr rt pcr kit, by Takara Bio (1 mentions)
Kapataq extra hotstart readymix with dye, by Roche (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Rt master mix, by Toyobo (1 mentions)
Sybr green 1 master mix, by Roche (1 mentions)
Farb buffer, by Favorgen Biotech (1 mentions)
Sybr green pcr master mix, by Roche (1 mentions)
Quantstudio 1 real time pcr system, by Thermo Fisher Scientific (1 mentions)
7 protocols using blood cultured cell total rna mini kit
1
Quantification of Survivin mRNA Expression
2
Quantifying S. aureus RNA Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total S. aureus RNA was isolated using a Blood / Cultured Cell Total RNA Mini Kit (Favorgen Biotech Corp., Ping-Tung, Taiwan). Overnight cultures (4 × 103 CFU/ml) of the tested strains were inoculated into MHB in the presence or absence of Tokiinshi (1 to 5 mg/ml) and incubated with shaking for 10 h. Real-time qRT-PCR was performed using the cDNA prepared by ReverTra Ace (TOYOBO Co., Ltd., Osaka, Tokyo). Primers designed for the qRT-PCR assays are listed in S1 Table [24 (link)]. All samples were analyzed in triplicate, and expression levels normalized against gmk gene expression [25 (link)]. The results are shown as the mean ± SE, which were derived from at least three independent experiments.
3
RNA Extraction and RT-PCR Analysis of PC12 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA isolation from differentiated PC12 cells was performed using a Blood/Cultured Cell Total RNA mini kit (Favorgen Biotech Corp., Taiwan). The extracted total RNA was subjected to reverse transcriptase reaction using a PrimeScript real-time PCR (RT-PCR) kit (TAKARA, Osaka, Japan). PCR was performed using KAPATaq EXtra HotStart ReadyMix with dye (KAPA Biosystems Inc., Woburn, MA, USA) and the following PCR primers: rat TrkA, 5′-ATG CTC GTC AGG ACT TCC ATC G-3′ and 5′-TAG CCA CAG CCA GAA GCT GC-3′; rat TrkB, 5′-AAG TCC TCT ATG AAG ACT GGA CC-3′ and 5′-TGC CAA ACT TGG AAT GTC TCG CCA-3′; rat TrkC, 5′-CAG CCC AGA GCC TTT GCT AAG-3′ and 5′-GGC AAA GGA GAG CCA GAG CCA TT-3′; rat p75NTR, 5′-CGG AAT TCG GAG ACA TGT TCC ACA GGC-3′ and 5′-CCT TGG GAT CCA TCG ACC-3′.
4
Quantitative Real-Time PCR Analysis of Immune Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMC, OVSAHO and MCAS cells were lysed in 350 μL/well of FARB buffer (Favorgen) containing 3.5 μL/well of 98% 2‐mercaptoethanol (Sigma‐Aldrich), and the Blood/Cultured Cell Total RNA Mini Kit (Favorgen) was used to extract total RNA from the cells. cDNA was synthesized using 5 × RT Master Mix (Toyobo) according to the manufacturer's instructions, and qRT‐PCR was performed in duplicate using SYBR Green I Master Mix and a LightCycler 480 (Roche) according to the manufacturer's protocols. Forward and reverse primer sequences were: PD‐L1, 5′‐GGCATTTGCTGAACGCAT‐3′ and 5′‐CAATTAGTGCAGCCAGGT‐3′; IL‐6, 5′‐ACAAGCCAGAGCTGTGCAGATG‐3′ and 5′‐GTGCCCATGCTACATTTGCCGA‐3′; TGF‐β, 5′‐GCTGCCTGTGTGACTTTGG‐3′ and 5′‐TCCTGGATTCTAGCACTTCTGG‐3′; ROR‐γt, 5′‐GTGGGGACAAGTCGTCTGG‐3′ and 5′‐AGTGCTGGCATCGGTTTCG‐3′; IFN‐γ, 5′‐CGAGGGTTGAATGAGAGCTT‐3′ and 5′‐CAGACGGCTGCCTTTATAGC‐3′; GAPDH, 5′‐GAAAGGTGAAGGTCGGAGTC‐3′ and 5′‐GAAGATGGTGATGGGATTTC‐3′; IL‐17, 5′‐AGAGATATCCCTCTGTGATC‐3′ and 5′‐CACCCCAAAATTGTCTCAGG‐3′; IL‐27, 5′‐GAGCAGCTCCCTGATGTTTC‐3′ and 5′‐AGCTGCATCCTCTCCATGTT‐3′; tumor necrosis factor α (TNF‐α), 5′‐GGCGTGGAGCTGAGAGATAAC‐3′ and 5′‐GGTGTGGGTGAGGAGCACAT‐3′; and IL‐23A, 5′‐CAGTTCTGCTTGCAAAGGAT‐3′ and 5′‐ATCTGCTGAGTCTCCCAGTG‐3′. All the Ab were from Sigma‐Aldrich. The ratio of each gene product to GAPDH was used to determine mRNA expression according to the cycle‐threshold method.
5
RNA Extraction from Cells and Supernatant
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted and purified from cells and culture supernatant using the Blood/Cultured Cell Total RNA Mini Kit (Favorgen Biotech, Pingtung City, Taiwan; No. FABRK 001–2) and the Viral Nucleic Acid Extraction Kit I (Favorgen Biotech; No. FAVNK 001–2), respectively, according to the manufacturer’s instructions.
6
Quantitative Analysis of Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from cells with a Blood/Cultured Cell Total RNA Mini Kit (FAVORGEN, Ping-Tung, Taiwan) according to the manufacturer's instructions. Total RNA was reverse-transcribed into cDNA by using ReverTra Ace® qPCR RT Master Mix (TOYOBO, Osaka, Japan). Quantitative real-time PCR was performed with SYBR Green PCR Master Mix (Roche Diagnostics KK, Tokyo, Japan) and the QuantStudio 1 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The expression levels of the target genes were normalized to β actin (internal control), and relative fold changes were calculated using the ΔΔCT method. The sequences of the oligonucleotides used as primers were as follows: sFlt-1 forward 5'-CTCCTGCGAAACCTCAGTG -3', reverse 5'-GACGATGGTGACGTTGATGT -3', PlGF forward 5'-GTTCAGCCCATCCTGTGTCT-3', reverse 5'-AACGTGCTGAGAGAACGTCA-3', VEGF forward 5'-TGCAGATTATGCGGATCAAACC-3', reverse 5'-TGCATTCACATTTGTTGTGCTGTAG-3', MMP (matrix metalloproteinase) 1 forward 5'-ACATGAGTCTTTGCCGGAGG-3', reverse 5'-ATCCCTTGCCTATCCAGGGT-3', MMP2 forward 5'-GCTGGCTGCCTTAGAACCTTTC-3', reverse 5'-GAACCATCACTATGTGGGCTGAGA-3', MMP9 forward 5'-GCCACTACTGTGCCTTTGAGTC-3', reverse 5'-CCCTCAGAGAATCGCCAGTACT-3', β actin forward 5'-CATGTACGTTGCTATCCAGGC-3', reverse 5'-CTCCTTAATGTCACGCACGAT-3'.
7
Quantifying Stress-Induced Transcriptional Changes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from live cells using a Blood/Cultured Cell Total RNA Mini Kit (FAVORGEN), followed by reverse transcription. cDNA was amplified for 40 cycles in a Light Cycler 480 (Roche). The expression of CHOP was normalized using GAPDH mRNA as an internal standard, and splicing of XBP1 (XBP1s/XBP1t) was calculated by the comparative Ct method.
The primer pairs used were as follows; human total XBP1 (XBP1t) 5′-GGCATCCTGGCTTGCCTCCA-3′ and 5′-GCCCCCTCAGCAGGTGTTCC-3′, human spliced XBP1 (XBP1s) 5′-CGCTTGGGGATGGATGCCCTG-3′ and 5′-CCTGCACCTGCTGCGGACT-3′, human CHOP 5′-GGAGCATCAGTCCCCCACTT-3′ and 5′-TGTGGGA TTGAGGGTCACATC-3′, and human GAPDH 5′-GAAA GGTGAAGGTCGGAGTC-3′ and 5′-GAAGATGGTGA TGGGATTTC-3′.
The primer pairs used were as follows; human total XBP1 (XBP1t) 5′-GGCATCCTGGCTTGCCTCCA-3′ and 5′-GCCCCCTCAGCAGGTGTTCC-3′, human spliced XBP1 (XBP1s) 5′-CGCTTGGGGATGGATGCCCTG-3′ and 5′-CCTGCACCTGCTGCGGACT-3′, human CHOP 5′-GGAGCATCAGTCCCCCACTT-3′ and 5′-TGTGGGA TTGAGGGTCACATC-3′, and human GAPDH 5′-GAAA GGTGAAGGTCGGAGTC-3′ and 5′-GAAGATGGTGA TGGGATTTC-3′.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!