Transcriptase 2

Manufactured by Thermo Fisher Scientific

Transcriptase II is a laboratory instrument designed for the reverse transcription of RNA into complementary DNA (cDNA). It performs the essential step of converting RNA into DNA, which is necessary for various molecular biology applications, such as gene expression analysis and RNA sequencing.

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Lab products found in correlation

Pcr primer, by Bioneer (4 mentions) Trizol reagent, by Thermo Fisher Scientific (3 mentions)

Spelling variants of this product

SuperScript II RNase H-Reverse Transcriptase (176 citations) SuperScript II reverse transcriptase and random primers (13 citations) SuperScript II Reverse Transcriptase reagents (5 citations) SuperScript II or III reverse transcriptase (4 citations) Superscript II RNase H2 reverse transcriptase kit (2 citations) Superscript(R) II reverse transcriptase (2 citations) SuperScript II reverse Transcriptase Mix (2 citations) SuperScript II RNase H-reverse transcriptase enzyme (2 citations) IMPROM II reverse transcriptase (2 citations) SuperScript II reverse transcriptase II (2 citations)

5 protocols using transcriptase 2

Quantifying Osteoblast Gene Expression

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Total RNA was isolated from cultured cells using TRIzol® reagent (Thermo Fisher Scientific. MO, USA), as stated in the manufacturer’s protocol. To quantify mRNA expression, total RNA (1 μg) was reverse transcribed and standard reverse transcription was carried out using Transcriptase II (both from Invitrogen). Reverse transcription polymerase chain reaction was performed using PCR primers (Bioneer, Daejeon, Korea) and annealing temperatures listed in Table 1. 18S rRNA was used as an endogenous control. The signal intensity of the product was normalized to the respective 18S rRNA signal intensity.Table 1

Osteoblast related marker primers used for RT-PCR

Gene name	Primer sequences(5'–3')	Annealing temperature (℃)	Product size (bp)
ALP	Forward: CCC ACG TTT TCA CAT TCG GT	57	190
ALP	Reverse: GCC TGG TAG TTG TTG TGA GC	57	190
BSP	Forward: TTT ATC CTC CTC TGA AAC GGT	55	110
BSP	Reverse: GTT TGA AGT CTC CTC TTC CTC C	55	110
OCN	Forward: GCG CTC TGT CTC TCT GAC CT	58	225
OCN	Reverse: TTT GTA GGC GGT CTT CAA GC	58	225
OPN	Forward: ACA CTT TCA CTC CAA TCG TCC	58	240
OPN	Reverse: TGC CCT TTC CGT TGT TGT CC	58	240
PINK1	Forward: CAC ACT GTT CCT CGT TAT GAA GA	56	157
PINK1	Reverse: CTT GAG ATC CCG ATG GGC AAT	56	157
18S	Forward: CTG AGA AAC GGC TAC CAC ATC	58	107
18S	Reverse: GCC TCG AAA GAG TCC TGT ATT G	58	107

ALP alkaline phosphatase, BSP bone sialoprotein, OCN osteocalcin; OPN osteopontin, PINK1 PTEN-induced kinase 1

Lee S.Y., An H.J., Kim J.M., Sung M.J., Kim D.K., Kim H.K., Oh J., Jeong H.Y., Lee Y.H., Yang T., Kim J.H., Lim H.J, & Lee S. (2021). PINK1 deficiency impairs osteoblast differentiation through aberrant mitochondrial homeostasis. Stem Cell Research & Therapy, 12, 589.

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RNA Extraction and RT-PCR Analysis

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Total RNA was extracted using TRIzol reagent, and standard reverse transcription (RT) was carried out using transcriptase II (both from Invitrogen). RT-PCR was performed using PCR primers (Bioneer, Daejeon, Korea) and annealing temperatures listed in Table I. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. The signal intensity of the product was normalized to the respective GAPDH signal intensity. Osteoinductive medium-treated cells were used as control.

Heo J.S., Lee S.G, & Kim H.O. (2017). Distal-less homeobox 5 is a master regulator of the osteogenesis of human mesenchymal stem cells. International Journal of Molecular Medicine, 40(5), 1486-1494.

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Quantitative RT-PCR Analysis of hiPS Cells

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Total RNA was extracted using TRIzol reagent (Invitrogen). Standard reverse transcription (RT) was performed using transcriptase II (Invitrogen). RT-PCR was performed using PCR primers (Bioneer, Daejeon, Korea) under the conditions listed in Table I. The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) level was used as an internal control. Human induced pluripotent stem (hiPS) cell cDNA was used as a positive control (kindly provided by Dr Dong-Wook Kim, Yonsei University College of Medicine). The signal intensity of the product was normalized to its respective GAPDH signal intensity.

HEO J.S., CHOI Y., KIM H.S, & KIM H.O. (2015). Comparison of molecular profiles of human mesenchymal stem cells derived from bone marrow, umbilical cord blood, placenta and adipose tissue. International Journal of Molecular Medicine, 37(1), 115-125.

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Total RNA Isolation and RT-PCR Analysis

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Total RNA was isolated using TRIzol reagent (Invitrogen). Standard reverse transcription was performed using transcriptase II (Invitrogen). RT-PCR was performed using PCR primers (Bioneer, Daejeon, Korea) under the conditions listed in Table 1. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) level was used as an internal control. The signal intensity of the product was normalized to its respective GAPDH signal intensity.

Heo J.S., Lee S.G, & Kim H.O. (2017). The Flavonoid Glabridin Induces OCT4 to Enhance Osteogenetic Potential in Mesenchymal Stem Cells. Stem Cells International, 2017, 6921703.

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Quantitative Gene Expression Analysis

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Total RNA was prepared using TRIzol reagent, and cDNA was synthesized using transcriptase II (Invitrogen). RT-PCR was performed with PCR primers under the conditions listed in Table 1 (Bioneer). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal standard. The signal intensity of the product was normalized to its respective GAPDH signal intensity.

Heo J.S., Kim H.O., Song S.Y., Lew D.H., Choi Y, & Kim S. (2016). Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo. BioMed Research International, 2016, 8196078.

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