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Thermal cycle dice tp800

Manufactured by Takara Bio
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The Thermal Cycle DiceTM TP800 is a laboratory instrument designed for thermal cycling applications. It is capable of precisely controlling and regulating temperature changes to support various scientific procedures.

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Lab products found in correlation

Sybr premix ex taq, by Takara Bio (2 mentions) Isogen kit, by Nippon Gene (1 mentions) Superscript 3 first strand synthesis system for rt pcr, by Thermo Fisher Scientific (1 mentions) Sybr premix ex taqtm 2 tli rnaseh plus, by Takara Bio (1 mentions) Rq1 rnase free dnase, by Promega (1 mentions) Revatra ace, by Toyobo (1 mentions) Primescript 1st strand cdna synthesis kit, by Takara Bio (1 mentions) Fastpure rna kit, by Takara Bio (1 mentions) Miscript sybr green pcr kit, by Qiagen (1 mentions) Miscript reverse transcription kit, by Qiagen (1 mentions) Mem per plus kit, by Thermo Fisher Scientific (1 mentions) Proteojet, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Primescript rt master mix, by Takara Bio (1 mentions) Rnase free dnase set, by Qiagen (1 mentions)

Close spelling variants of this product

TP800 (16 citations) Thermal Cycle Dice Real Time system TP800 (4 citations) Thermal Cycle Dice (3 citations)

6 protocols using thermal cycle dice tp800

1

Quantification of Retrotransposon Expression

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Expression levels of retrotransposons were evaluated by RT-qPCR. Total testicular RNA (1 μg) was isolated using an ISOGEN kit (Nippon Gene, Tokyo, Japan), treated with RQ1 RNase-free DNase (Promega, Madison, WI, USA),
and reverse-transcribed in the presence of random hexamers using a Superscript III first-strand synthesis system for RT-PCR (Thermo Fischer Scientific). An aliquot of synthesized cDNA was subjected to PCR using the SYBR
Premix Ex TaqTM II (Tli RNase H Plus) (TAKARA Bio, Otsu, Shiga, Japan) in a Thermal Cycle DiceTM TP800 (TAKARA Bio). The mRNA levels were normalized to β-actin (Actb) mRNA levels.
Data are presented as mean ± SEM (n ≥ 3). Student’s t-tests were used for statistical analyses; significance was assumed at P < 0.05. Primer sequences used for PCRs are: Actb,
5’-AGATCAAGATCATTGCTCCTCCT-3’ (sense) and 5’-ACGCAGCTCAGTAACAGTCC-3’ (antisense); IAP (Intracisternal A-particle element), 5’-AACCAATGCTAATTTCACCTTGGT-3’ (sense) and 5’-GCCAATCAGCAGGCGTTAGT-3’ (antisense); LINE-1 (Long
interspersed element-1), 5’-GGCGAAAGGCAAACGTAAGA-3’ (sense) and 5’-GGAGTGCTGCGTTCTGATGA-3’ (antisense); SINE B1 (Short interspersed element B1), 5’-TGAGTTCGAGGCCAGCCTGGTCTA-3’ (sense) and 5’-ACAGGGTTTCTCTGTGTAGCCCTG-3’
(antisense).
KASHIWABARA S.I., TSURUTA S., YAMAOKA Y., OYAMA K., IWAZAKI C, & BABA T. (2017). PAPOLB/TPAP regulates spermiogenesis independently of chromatoid body-associated factors. The Journal of Reproduction and Development, 64(1), 25-31.
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2

RNA Extraction and qRT-PCR Analysis

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Tissues and cells were homogenized in 1 mL of Sepazol and total RNA was extracted according to the manufacturer's instructions (Nacalai tesque). cDNA was synthesized from total RNA using RevaTraAce reverse transcriptase (Toyobo) and oligo dT primer. Real-time PCRs were performed to amplify fragments representing for the indicated mRNA expression using the Thermal Cycle DiceTM TP800 (Takara) and SYBR Premix Ex Taq (Takara). The primer sequences can be found in Supplementary Table S1.
Hiyoshi H., Goto N., Tsuchiya M., Iida K., Nakajima Y., Hirata N., Kanda Y., Nagasawa K, & Yanagisawa J. (2014). 2-(4-Hydroxy-3-methoxyphenyl)-benzothiazole suppresses tumor progression and metastatic potential of breast cancer cells by inducing ubiquitin ligase CHIP. Scientific Reports, 4, 7095.
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3

Quantifying mRNA Expression via qRT-PCR

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Total RNA was isolated with a FastPure® RNA kit (Takara Bio, Shiga, Japan) according to the manufacturer’s instructions. Total RNA (1 μg) was reverse transcribed with the PrimeScript® 1st Strand cDNA Synthesis kit (Takara Bio). The real-time quantitative PCR analysis was performed to amplify fragments representing the indicated mRNA or for pre-rRNA expression using the Thermal Cycle DiceTM TP800 (Takara) and SYBR. Indicated mRNA or pre-rRNA levels were normalised to β-actin mRNA levels. The primer sequences can be found in Supplementary Table S2.
Kumazawa T., Nishimura K., Katagiri N., Hashimoto S., Hayashi Y, & Kimura K. (2015). Gradual reduction in rRNA transcription triggers p53 acetylation and apoptosis via MYBBP1A. Scientific Reports, 5, 10854.
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4

Protein and miRNA Analysis Methods

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Tissue homogenisation and western blotting analysis were performed using conventional methods14 (link),31 (link),32 (link). Isolation of membrane and cytoplasmic protein fractions were performed following the manufacturer’s protocols (Fermentas, ProteoJET; Thermo, Mem-PER Plus kit). RNA purification, reverse transcription reaction, and real-time qPCR analysis (Takara Bio, Thermal Cycle Dice (TP800), Thermal Cycle Dice Real Time System Ver. 5. 11B) were performed by conventional methods14 (link),31 (link),32 (link). Briefly, to detect miRNAs, cDNA was generated using an miScriptReverse Transcription Kit (QIAGEN). Expression of miR-16, miR-23 and miR-21 was detected using an miScript SYBR Green PCR Kit (QIAGEN) with the following primers: 5'- TAGCAGCACGTAAATATTGG-3' for miR-16, 5'-ATCACATTGCCAGGGATTTCC-3' for miR-23 and 5'-TAGCTTATCAGACTGATGTTGA-3' for miR-21.
Ageta H., Ageta-Ishihara N., Hitachi K., Karayel O., Onouchi T., Yamaguchi H., Kahyo T., Hatanaka K., Ikegami K., Yoshioka Y., Nakamura K., Kosaka N., Nakatani M., Uezumi A., Ide T., Tsutsumi Y., Sugimura H., Kinoshita M., Ochiya T., Mann M., Setou M, & Tsuchida K. (2018). UBL3 modification influences protein sorting to small extracellular vesicles. Nature Communications, 9, 3936.
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5

Real-Time PCR Quantification Protocol

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Real-time PCR was carried out using Thermal Cycle Dice TP800 (TaKaRa) and SYBR Premix Ex Taq (TaKaRa). A PCR mixture with a total volume of 20 µL containing 1 µL of 10 µM primer for each and 2 µL of indicated template DNA solution was prepared in accordance with the manufacturer's instructions. The PCR reaction was started at 95°C, followed by 40 cycles of dissociation at 94°C for 5 s and annealing and elongation at 60°C for 30 s.
Kitamura R., Miura N., Okada K., Motone K., Takagi T., Ueda M, & Kataoka M. (2020). Design of novel primer sets for easy detection of Ruegeria species from seawater. Bioscience, biotechnology, and biochemistry, 84(4).
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6

RNA Extraction and qPCR Analysis of O. minor

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Total RNA was isolated from germinating seeds of O. minor using TRIzol Reagent (Thermo Fischer Scientific, Waltham, MA, USA) and treated with the RNase-Free DNase Set (Qiagen, Hilden, Germany) in accordance with the manufacturer's instructions. Synthesis of cDNA and quantitative PCR was conducted using the PrimeScript RT Master Mix (Takara Bio, Kusatsu, Japan) with Thermal Cycle Dice TP800 (Takara Bio). The sequences of gene-specific primers used were as follows:
Rv, 5′-CGATGGGAATTCAGACGACA-3′. The gene comp71446_c0_seq1 was shown to be constitutively expressed in our previous transcriptome analysis (Okazawa et al, 2020) , and was selected as a reference gene for normalization using the 2 -∆Ct method.
Okazawa A., Baba A., Okano H., Tokunaga T., Nakaue T., Ogawa T., Shimma S., Sugimoto Y., & Ohta D. (2021). Localization of planteose hydrolysis during seed germination of Orobanche minor.
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