Total RNA was isolated from the abdominal cells using TRIzol (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. The quality assessment of the RNA and synthesized cDNA was performed according to our previously published method (13 (link)). The PCR amplification of the P2X7 receptor and β-actin (internal standard for quantification) genes was performed according to our previously published method (20 (link)). The amplification system included 2 µl cDNA, 12.5 µl PCR mixture (Tiangen Biotech Co., Ltd.), 2 µl primers (1 µl each of sense and antisense primers) and 8.5 µl nuclease-free water. The sequences of the primers used for RT-PCR analysis were as follows: P2X7 receptor (171 bp): sense, 5'-GCACGAATTATGGCACCGTC-3' and antisense, 5'-CCCCACCCTCTGTGACATTC-3'; uc.48+ (231 bp): sense, 5'-GCAAACTGGATGAGGAT-3' and antisense, 5'-GTAGTGCCACAAGGAGA-3'; β-actin (240 bp): sense, 5'-TAAAGACCTCTATGCCAACACAGT-3' and antisense, 5'-CACGATGGAGGGGCCGGACTCATC-3'. The PCR conditions used to detect these genes and analyze the PCR products were as described in the study by Wu et al (13 (link)).
Pcr mixture
Manufactured by Tiangen Biotech
Sourced in China
The PCR mixture is a reagent solution used in the Polymerase Chain Reaction (PCR) process. It contains the necessary components for DNA amplification, including DNA polymerase, deoxyribonucleotides (dNTPs), and buffer solution. The PCR mixture enables the exponential replication of specific DNA sequences, a fundamental technique in molecular biology and genetics.
Automatically generated - may contain errors
Lab products found in correlation
Trizol, by Thermo Fisher Scientific (1 mentions)
Primescript rt master mix perfect real time kit, by Takara Bio (1 mentions)
Reverse transcription kit, by Tiangen Biotech (1 mentions)
Streptozocin stz, by MultiSciences Biotech (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Rosiglitazone, by MultiSciences Biotech (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Cyt c, by Cell Signaling Technology (1 mentions)
3 protocols using pcr mixture
1
Quantification of P2X7 Receptor Expression
2
Quantitative RT-PCR for Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was prepared as mentioned above using the same samples. PrimeScript RT Master Mix Perfect Real Time kit (Takara) was used to synthesize first strand cDNA as described by the manufacturer. Specific DNA sequences were amplified with a PCR mixture (TIANGEN Biotech, China) and resolved on a 2% agarose gel. PCR primers were indicated as follows: Apaf 1, 5′-ATGTTATCCCTGTGGAGAG- TGG-3′ (sense) and 5′-CACCAACTAAAGACACGACGAG-3′ (antisense); Bace 2, 5′-TTGTGGACACCGGAAGCAGTAA-3′ (sense) and 5′-CCTCAAAGCCCTTGGAGTGGTA-3′ (antisense); Plcb 4, 5′-GCCCATTACTTCATCAGTTCCT-3′ (sense), 5′-TACACATTGCTTTTCCGTGAGT-3′ (antisense); β-actin, 5′-CACCCGCGAGTACAACC TTC-3′ (sense) and 5′-CCCATACCCACCATCACACC-3′ (antisense).
3
Establishing Diabetic Cardiomyopathy Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The MGE IIe (purity 98%) was purchased from Honghe Qianshan Bioengineering Co., Ltd. (Honghe, China). Streptozocin (STZ) and rosiglitazone were obtained from Multi Sciences Biotech Co. Ltd. (Hangzhou, China). Antibodies against ß-actin, caspase-3, -8, -9, -12, Bax, Cyt-C, and Bcl-2 were obtained from Cell Signaling Technology (Danvers, MA, United States). ELISA kits for the detection of IL-1, IL-6, TNF-α, CK-MB, HDL, and LDL were purchased from Thermo Fisher Scientific (Waltham, MA, United States). The mRNA extraction kit, reverse transcription kit, PCR mixture, and Masson staining kit were purchased from Tiangen Biochemical Technology Co., Ltd. (Beijing, China). The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies (Kumamoto, Japan).
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!