For mtDNA, total DNA was isolated using a Total DNA Extraction Kit (TIANGEN, Beijing, China). Relative levels of mtDNA copy number were determined by real-time PCR. The reactions were performed on a LightCycler® 96 Real-Time PCR System (Roche). The data are displayed as 2-ΔΔCt values and are representative of at least three independent experiments. The following primers sets were used: Mt-Mito Forward: 5′-CACTTTCCACACAGACATCA-3′, Reverse: 5′-TGGTTAGGCTGGTGTTAGGG-3′; B2M Forward: 5′-TGTTCCTGCTGGGTAGCTCT-3′, Reverse: 5′-CCTCCATGATGCTGCTTACA-3′. To determine mitochondrial mass, cells were stained with 100 nM MitoTracker Red (Invitrogen) at 37°C for 15 min and then analyzed by flow cytometry. The relative fluorescence intensity was record and represented as mitochondrial mass.
Total dna extraction kit
Manufactured by Tiangen Biotech
Sourced in China
The Total DNA Extraction Kit is a laboratory equipment designed to extract high-quality DNA from a variety of biological samples. It employs a simple and efficient protocol to isolate genomic DNA, enabling researchers to obtain pure DNA samples for downstream applications such as PCR, sequencing, and molecular analysis.
Automatically generated - may contain errors
Lab products found in correlation
Gel extraction kit, by Tiangen Biotech (2 mentions)
Pcr purification kit, by Tiangen Biotech (2 mentions)
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions)
T4 dna ligase, by Thermo Fisher Scientific (2 mentions)
Dnase 1, by Thermo Fisher Scientific (2 mentions)
Mitotracker red, by Thermo Fisher Scientific (1 mentions)
Lightcycler 96 real time pcr system, by Roche (1 mentions)
Primer premier software, by Premier Biosoft (1 mentions)
Cfx96 real time pcr system, by Bio-Rad (1 mentions)
Ionomycin, by Solarbio (1 mentions)
Anti cd4 apc, by BD (1 mentions)
Fitc anti il 17a, by BD (1 mentions)
Anti cd25 pe, by BD (1 mentions)
Metronidazole, by Solarbio (1 mentions)
Total rna extraction kit, by Tiangen Biotech (1 mentions)
Bicinchoninic acid (bca), by Nanjing Jiancheng (1 mentions)
Ciprofloxacin, by Solarbio (1 mentions)
Chloroform, by Thermo Fisher Scientific (1 mentions)
Primuline, by Merck Group (1 mentions)
Plasmid miniprep kit, by Tiangen Biotech (1 mentions)
6 protocols using total dna extraction kit
1
Quantify Mitochondrial DNA and Mass
2
Telomere Length Quantification by qPCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The average telomere length was measured using a real-time PCR assay as previously described (Gil and Coetzer, 2004 (link)). Briefly, telomere length was quantified by the ratio of the amount of the telomere amplification product (T) to the amount of product from a single-copy gene (S). The single-copy gene 36B4 was used as the control. The PCR reaction was performed on a CFX96 real-time PCR system (Bio-Rad). The genomic template DNA was extracted using a Total DNA Extraction Kit (Tiangen Biotech Co. Ltd.). The primers used to amplify the telomere DNA and the control gene 36B4 were designed using Primer Premier software (Premier Biosoft Inc., CA) and are shown in Table1 . The conditions for the PCR reaction were 95°C for 5 min and 40 cycles of 95°C for 10 s, 60°C for 30 s. The telomere DNA was normalized to that of 36B4 to generate a T/S ratio which indicates the relative length of the telomere.
3
Berberine Impacts on Inflammatory Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The standard of berberine (C20H18NO4, molecular weight: 235.324 kD) was obtained from Solarbio Biotechnology Co., Ltd. (Beijing, China). Dextran sulfate sodium (DSS molecular weight: 36000-50000kD), ciprofloxacin, metronidazole, Phorbol 12-myristate (PMA), ionomycin, and Bravertin A were also obtained from Solarbio Biotechnology Co., Ltd. (Beijing, China). Total DNA extraction kit, total RNA extraction kit, first-stand cDNA reverse transcription kit, polymerase chain reaction kit, and primers were obtained from TianGen Biotechnology Co., Ltd. (Beijing, China). Mouse IL-10 and IL-17 ELISA kits were obtained from Multi Science Biotechnology Co., Ltd. (Hangzhou, China). APC anti-CD4, FITC anti-IL17A, PE anti-CD25, and Alexa Fluor 488 anti-Foxp3 antibodies for flow cytometry were purchased from BD bioscience Co., Ltd. (Franklin Lakes, NJ, United States). BCA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Cr) test kit were purchased from Nanjing Jiancheng Biological Engineering Institute (Nanjing, China).
4
Comprehensive Metabolite Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chemicals including primuline, hexane, acetone, acetic acid, and diethyl ester were purchased from Sigma-Aldrich (St. Louis, USA) or Sinopharm (Shanghai, China). Methanol, chloroform, Phusion polymerase, restriction enzymes, T4 DNA ligase, DNase I, and RevertAid First Strand cDNA Synthesis Kit were purchased from Thermo Fisher Scientific (Beverly, USA). Plasmid Miniprep Kit, Total DNA Extraction Kit, PCR Purification Kit, and Gel Extraction Kit were purchased from TianGen (Beijing, China). RNA Protect Reagent and RNeasy Mini Kit were purchased from Qiagen (Hilden, Germany). Power SYBR® Green PCR Master Mix was from Applied Biosystems (Massachusetts, USA). FFA, DAG, TAG, or FAEE standards were purchased from Nu-Chek (Elysian, USA) or Larodan (Stockholm, Sweden).
5
Molecular Cloning and Protein Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Restriction enzymes, polymerase, and ligase were purchased from New England Biolabs (Beverly, USA). The plasmid miniprep kit, total DNA extraction kit, PCR purification kit, and gel extraction kit were purchased from TianGen (Beijing, China). Primers and genes were synthesized by TsingKe (Beijing, China). Fatty acid standards were purchased from Larodan (Stockholm, Sweden). Chemical reagents were purchased from Sigma-Aldrich (St. Louis, USA) or Sinopharm (Shanghai, China). His-tag and S-tag antibodies were purchased from Proteintech (Wuhan, China).
6
Quantifying Mitochondrial DNA Copy Number
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ratio of the levels of the single-copy mitochondrial gene Cox1 (cytochrome c oxidase) to the single-copy nuclear gene Nthl1 (nth endonuclease III-like 1) was used to determine the mitochondrial DNA copy number as described in a previous study (Aiken et al., 2016 (link)). Real-time PCR was performed to obtain the average copy number of mitochondrial DNA/nuclear DNA. Total DNA was extracted using a Total DNA Extraction Kit (Tiangen Biotech Co. Ltd.), from 9 mice per group. Primer design and PCR conditions were the same as those used for telomere length measurement. The method for calculating the average mitochondrial DNA copy number followed that described in a previous study (Nicklas et al., 2004 (link)).
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!