Total RNA of cells was extracted using TRIzol Reagent (Invitrogen, Waltham, MA, USA). First-strand complementary DNA (cDNA) was synthesized from 500 ng total. RNA using the high-capacity cDNA reverse transcription kit (Applied Biosystems, Foster City, CA, USA). Quantitative polymerase chain reaction (PCR) was performed in a 10 μL standard PCR reaction mixture prepared in duplicate using an Applied Biosystems 7900 Prism Real-Time PCR system and SYBR Premix Ex Taq (TaKaRa, Dalian, Japan), in accordance to the manufacturer’s protocol. Quantitative PCR primers were as follows: BACE1, sense: 5′-gactagga-gaccagaagtgaatg-3′ and antisense: 5′-cttctgtccaccc-tattttctgg-3′. The β-actin primers were used as the internal control. β-actin, sense: 5′-cacagactacctcatgaagatcc-3′ and antisense: 5′-cagctcgtaactcttctccag −3′.
7900 prism real time pcr system
Manufactured by Takara Bio
Sourced in Japan
The 7900 Prism Real-Time PCR system is a high-performance real-time PCR instrument designed for accurate and reliable nucleic acid quantification. It features a thermal cycler, optical detection system, and software for data analysis.
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Lab products found in correlation
3 protocols using 7900 prism real time pcr system
1
Quantitative Analysis of BACE1 Expression
2
Quantitative PCR of Total RNA
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Total RNA from the cells was extracted by TRIzon Reagent method (CWBIO, WUHAN). First-strand complementary DNA (cDNA) was synthesized from a total of 50mg RNA using the ReverTra Ace qPCR RT kit (TOYOBO, USA). Quantitative polymerase chain reaction (PCR) was performed in a standard PCR reaction mixture prepared in duplicate using an Applied Biosystems 7900 Prism Real-Time PCR system and SYBR Premix Ex Taq (TaKaRa, Dalian, Japan) in accordance with the manufacturer’s protocol. Quantitative PCR primers were as follows: sense: 5′-CTGCAATGTGCCTTTTCTGA-3′, antisense: 5′-ATGCTTCCCTCAAGCATCTG-3′
3
Quantitative Analysis of BACE1 Expression
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