Total RNA was extracted from RCC cell lines and the clinical primary tissues, using the TRI Reagent and Tianamp Genomic DNA Kits (TIANGEN®, Shanghai, China) as described in our previous study.26 (link) Genomic DNA of primary RCC tissues and RCC cells was isolated using Tianamp Genomic DNA Kits (TIANGEN®, Shanghai, China) according to the manufacturer’s instructions. DNA and RNA concentrations were assessed using the ND2000 Spectrophotometer (Thermo Fisher Scientific).
Tianamp genomic dna kit
Manufactured by Tiangen Biotech
Sourced in China, Germany
The TIANamp Genomic DNA Kit is a DNA extraction kit designed to efficiently isolate genomic DNA from a variety of sample types. The kit utilizes a silica-based membrane technology to capture and purify DNA, while removing contaminants and inhibitors. The extracted DNA is suitable for use in various downstream applications, such as PCR, sequencing, and other molecular biology techniques.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 2000, by Thermo Fisher Scientific (45 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (35 mentions)
Trizol reagent, by Thermo Fisher Scientific (25 mentions)
Ez dna methylation gold kit, by Zymo Research (23 mentions)
Epitect bisulfite kit, by Qiagen (20 mentions)
Nanodrop, by Thermo Fisher Scientific (18 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (14 mentions)
Ez dna methylation kit, by Zymo Research (13 mentions)
Pgm t vector, by Tiangen Biotech (12 mentions)
Pmd19 t vector, by Takara Bio (10 mentions)
Trizol, by Thermo Fisher Scientific (10 mentions)
Nanodrop 2000c spectrophotometer, by Thermo Fisher Scientific (9 mentions)
Universal dna purification kit, by Tiangen Biotech (9 mentions)
Dna bisulfite conversion kit, by Tiangen Biotech (8 mentions)
Pmd18 t vector, by Takara Bio (8 mentions)
Nanodrop spectrophotometer, by Thermo Fisher Scientific (8 mentions)
Qubit dsdna hs assay kit, by Thermo Fisher Scientific (8 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Novaseq 6000, by Illumina (6 mentions)
Hiseq 2500, by Illumina (6 mentions)
1 032 protocols using tianamp genomic dna kit
1
RNA and DNA Extraction from RCC Cells
2
Characterization of iPSC-derived Lineages
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Following 14 days of incubation as described above, genomic DNA from iPSCs and EBs was extracted using a TIANamp Genomic DNA kit (Tiangen Biotech Co., Ltd., Beijing, China) according to the manufacturer's protocol. PCR was used to examine the expression of genes representative of the endoderm, mesoderm and ectoderm. The extracted genomic DNA of iPSCs and EBs was mixed with primers and a TIANamp Genomic DNA kit (Tiangen Biotech Co., Ltd.) and the thermocycling conditions were as follows: Pre-denaturation at 94°C for 4 min, denaturation at 94°C for 30 sec, annealing at 55°C for 30 sec and extension at 72°C for 30 sec for 35 cycles, followed by a final elongation at 72°C for 2 min and storage at 4°C. The amplified PCR products were resolved on 1.5% agarose gels (Thermo Fisher Scientific, Inc.) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. The gels were run for 25 min at 100 V. Images were captured using a Bio-Rad Gel document system (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The primer sequences of all primers are listed in Table I .
3
Detection of Vibrio alginolyticus in Shrimp Samples
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Shrimp were purchased from a local market and verified to be free of V. alginolyticus by PCR [40 (link)]. The PCR primers are listed in Table 1 . To test the detection limit in the spiked shrimp sample, 1 mL of serially diluted V. alginolyticus culture from 2.5 × 104 CFU/mL to 2.5 CFU/mL was mixed with 1 g of shrimp homogenate (shrimp were ground in liquid nitrogen) and 9 mL alkaline peptone broth (Sinopharm Chemical Reagent Co., Ltd., China). Genomic DNA was purified using the TIANamp Genomic DNA Kit (Tiangen Biotech) in a 50 μL volume. One microliter of the purified DNA was used for RPA-LFD. For application simulation, the shrimp was cut into small pieces, and 300 mg of the pieces was ground in liquid nitrogen for each sample. Into several randomly selected homogenate samples, 3 μL of the 106 CFU/mL V. alginolyticus culture was mixed. DNA was purified from the homogenate samples by the TIANamp Genomic DNA Kit (Tiangen Biotech) in a 50 μL volume. One microliter of the purified DNA was used for RPA-LFD or PCR detection.
4
DNA Extraction from Cell Lines and Pig Breeds
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Genomic DNA of 3D4/21 cells was extracted by the TIANamp Genomic DNA kit (TIANGEN, Beijing, China). Genomic DNA of Duroc, Meishan, and Yorkshire pigs were extracted from the blood sample by TIANamp Genomic DNA kit (TIANGEN, Beijing, China).
5
Biodistribution of Oncolytic Adenovirus
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Huh7 tumor-bearing nude mice received a dose of 109 OV intratumorally. Organs and tissue samples were collected and snap frozen in liquid nitrogen at day 7 or day 14 after administration. Adenoviral genome in tissue lysate was extracted using TIANamp Genomic DNA Kit (Tiangen Biotech) after homogenization. Viral titers from individual organ and tissue samples were determined by qPCR (Supplementary Fig. 4 ). HepG2 tumor-bearing mice received a dose of 109 OV intratumorally. Organs and tissue samples were collected and snap frozen in liquid nitrogen at day 7 after administration. Adenoviral DNA and RNA in tissue lysate were extracted using TIANamp Genomic DNA Kit (Tiangen Biotech) and RNAiso Plus (TaKaR). Viral titers and relative RNA expression level from individual organ and tissue samples were determined by qPCR with qL2 primers and GAPDH primers (Fig. 5c , Supplementary Fig. 2a, b ).
6
Genotyping PARG Knockout Mice
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Genomic DNA was purified from mouse tails using TianAMP genomic DNA kits (Tiangen, Beijing, China). The concentration and the quality of DNA were assessed by ultraviolet (UV) absorbance using a NanoDrop ND-2000 spectrophotometer (Thermo Fisher Scientific). The DNA was then amplified by PCR (94°C for 2 min; 10 cycles of 94°C for 20 s, 65°C for 15 s, and 68°C for 10 s; 10 cycles of 94°C for 15 s, 60°C for 15 s, and 72°C for 10 s; 72°C for 2 min, 10°C hold) using primers provided by the Jackson Laboratory (Wild-type Forward: 5′-GAG ATA TCT AAG TCA GAG AAA GGT GGT-3′, Wild-type Reverse: 5′-CCT CCT CTG GTG TGT CTG AAG-3′, Mutant Forward: 5′-CGG TCG CTA CCA TTA CCA GT-3′, Mutant Reverse: 5′-GGT ATC AGC GAT GGT TGT TC-3′). The PCR products were 279 bp for the WT sample, and 279 and 507 bp for the heterozygous PARG knockout (PARG+/−) sample.
7
Isolation and Characterization of CD133+ Hep-2 Cells
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Hep-2 cells were purchased from the Cell Research Institute of the Chinese Academy of Sciences (Shanghai, China). RPMI-1640 medium was obtained from Gibco-BRL (Gaithersburg, MD, USA). Fetal bovine serum was obtained from Hyclone (Logan, UT, USA). CD133 MicroBeads Kits were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). Anti-mouse CD133-PE antibody was purchased from Abcam (Cambridge, MA, USA). Cell Counting Kit-8 (CCK-8) kits were purchased from 7Sea Biotech (Shanghai, China). Annexin V kits were purchased from KeyGEN BioTECH (Jiangshu, China). TIANamp Genomic DNA Kits were purchased from Tiangen Biotech (Beijing, China). REeverAid™ first strand cDNA synthesis kits were purchased from Toyobo (Osaka, Japan). SYBR Green PCR kits were purchased from Thermo Scientific (Waltham, MA, USA). WesternBright™ ECL was purchased from Advansta (San Jose, CA, USA). Matrigel and Transwell chambers were purchased from Costar (Temecula, CA, USA). Anti-GLUT-1, anti-RAD5, anti-PKcs antibody was purchased from Abcam. GAPDH was purchased from Cell Signaling Technology (Beverly, MA, USA). Healthy male NOD/SCID nude mice (aged 4–6 weeks), weighing 18–20 g, were provided by Shanghai Sleek Laboratory Animal Co. Ltd. (Shanghai, China).
8
RNA and DNA Extraction from RCC Tissues
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Total RNA and genomic DNA of primary RCC tissues and cell lines were extracted using an RNA-easy Isolation Reagent (Vazyme Biotech, Nanjing, China) and TIANamp Genomic DNA Kits (TIANGEN, Shanghai, China) according to their instructions, respectively, as previously described (Zhang et al., 2014 (link); Wang et al., 2019 (link)).
9
Aphid DNA Extraction Protocol
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Prior to DNA extractions, each aphid sample was washed for 5 min in 70% ethanol and rinsed three times with sterile water in 2.0 ml Eppendorf tubes to remove surface contaminants. TIANamp Genomic DNA Kits (TIANGEN Biotech (Beijing) LTD., China) were used for DNA extractions. The protocol that was followed has been previously reported (Zhao et al., 2016 ). The quantity and quality of the DNA were measured with a NanoDrop 2000C spectrophotometer (Thermo Scientific, USA).
10
Genomic DNA Extraction and Sequencing
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Genomic DNA from ear tissues was extracted using TIANamp Genomic DNA kits (Tiangen Biotech, Beijing, China). The concentration and purity of genomic DNA were detected using a NanoDrop™ 2000 (Thermo Fisher, Waltham, MA, USA). DNA samples with a light absorption ratio (A260/280) between 1.8 and 2.0 and a concentration > 50 ng/μL were used in the subsequent steps. DNA libraries were constructed for each individual using an MGIEasy FS DNA Prep kit (BGI, Shenzhen, China) following the manufacturer’s instructions. Paired-end sequencing using the MGISEQ-2000 platform (BGI, Shenzhen, China) yielded 150 bp-sized sequencing reads.
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