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Cell to ct lysis buffer

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The Cell-to-Ct lysis buffer is a reagent used for the direct lysis of cells in preparation for gene expression analysis. It is designed to facilitate the rapid and efficient extraction of RNA from cells, enabling direct downstream applications such as real-time PCR without the need for a separate RNA isolation step.

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Lab products found in correlation

Taqman gene expression assay, by Thermo Fisher Scientific (2 mentions) Cell to ct kit, by Thermo Fisher Scientific (2 mentions) 7500 real time pcr system, by Thermo Fisher Scientific (2 mentions) Primer express, by Thermo Fisher Scientific (1 mentions) 7900 real time pcr system, by Thermo Fisher Scientific (1 mentions) Collagenase hyaluronidase, by STEMCELL (1 mentions) Dnase 1, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Lysis buffer (523 citations) Cell lysis buffer (171 citations)

6 protocols using cell to ct lysis buffer

1

Cell Lysis and Gene Expression Analysis

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5 × 104 were lysed with 50 μl of Cell-to-Ct lysis buffer (Thermo Fisher). cDNA was directly synthesized from cell lysates with the Cell-to-Ct Kit (Thermo Fisher). Quantitative real-time PCR was performed with TaqMan Gene Expression Assays (Applied Biosystems) on a 7500 Real-Time PCR System (Applied Biosystems). β-actin was used as housekeeping gene.
Caielli S., Troggian Veiga D., Balasubramanian P., Athale S., Domic B., Murat E., Banchereau R., Xu Z., Chandra M., Chung C.H., Walters L., Baisch J., Wright T., Punaro M., Nassi L., Stewart K., Fuller J., Ucar D., Ueno H., Zhou J., Banchereau J, & Pascual V. (2018). A CD4+ T cell population expanded in Lupus blood provides B cell help through IL10 and succinate. Nature medicine, 25(1), 75-81.
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2

Cell Lysis and Gene Expression Analysis

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5 × 104 were lysed with 50 μl of Cell-to-Ct lysis buffer (Thermo Fisher). cDNA was directly synthesized from cell lysates with the Cell-to-Ct Kit (Thermo Fisher). Quantitative real-time PCR was performed with TaqMan Gene Expression Assays (Applied Biosystems) on a 7500 Real-Time PCR System (Applied Biosystems). β-actin was used as housekeeping gene.
Caielli S., Troggian Veiga D., Balasubramanian P., Athale S., Domic B., Murat E., Banchereau R., Xu Z., Chandra M., Chung C.H., Walters L., Baisch J., Wright T., Punaro M., Nassi L., Stewart K., Fuller J., Ucar D., Ueno H., Zhou J., Banchereau J, & Pascual V. (2018). A CD4+ T cell population expanded in Lupus blood provides B cell help through IL10 and succinate. Nature medicine, 25(1), 75-81.
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3

Transcriptome Analysis of Cultured Cumulus Cells

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After oocyte retrieval, the cumulus cells were mechanically stripped from the oocyte and isolated. Corona cells were removed with hyaluronidase, and mature oocytes were fertilized via intracytoplasmic sperm injection. The cumulus cells associated with these mature oocytes were lysed using cellto-CT lysis buffer (Thermo Fisher Scientific). Deoxyribonucleic acid (DNA) was removed by DNase treatment, and the resulting 30 mL lysate was stored at À80 C. Embryos were individually cultured to maintain embryo identity, ensuring that the cumulus cell transcriptome for a particular oocyte would be appropriately correlated with the pregnancy outcome for its associated embryo. Embryos were assessed on day 5 and 6 and graded using the blastocyst scoring system developed by Gardner and Schoolcraft (31) . All embryos in this analysis underwent trophectoderm biopsy for PGS at the blastocyst stage on day 6 prior to vitrification.
Green K.A., Franasiak J.M., Werner M.D., Tao X., Landis J.N., Scott RT J.r, & Treff N.R. (2018). Cumulus cell transcriptome profiling is not predictive of live birth after in vitro fertilization: a paired analysis of euploid sibling blastocysts. Fertility and sterility, 109(3).
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4

Isolation and Analysis of Mammary Gland Nuclei

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Mammary glands from pellet-bearing mice were harvest and digested with Collagenase/Hyaluronidase (Stem Cell Technologies). Digested tissue was further treated with 5X Tripsin (Life Technologies) for 5 minutes. Nuclei were isolated using sucrose gradient (Yamaji et al., 2013 (link)) and lysed with 30μl of Cell-To-Ct lysis buffer (Ambion). cDNA synthesis was performed according to manufacturer’s instructions. Real-time PCR were performed on a 7900 Real-Time PCR System (Applied Biosystems). Gene-specific primers were designed using Primer Express (Applied Biosystems) and qPCR reactions were performed using SYBR Green. Gapdh mRNA was used as endogenous control.
dos Santos C.O., Dolzhenko E., Hodges E., Smith A.D, & Hannon G.J. (2015). An epigenetic memory of pregnancy in the mouse mammary gland. Cell reports, 11(7), 1102-1109.
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5

Single-cell transcriptomics of cardiomyocytes

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Freshly isolated adult (~3-month old) cardiomyocytes and mCPCs (derived from GFP-cardiomyocytes) were captured using microfludic chip (Fig. 1f)41 (link)108 (link). The adult cardiomyocytes were used as controls. Briefly, isolated adult cardiomyocytes were preserved in EGTA-free KB solution9 (link), while mCPCs were kept in cold PBS during microfludic separation. Single-cells were isolated by encapsulation in droplet using a microfluidic device as described previously108 (link)109 (link). Once individual cells were isolated and verified under microscope, each was lysed with 2 μl of a customized buffer for whole genomic amplification for transcriptomic profiling. Alternatively, 5 μl Cell-to-Ct lysis buffer supplemented with DNase I (Ambion) was used to lyse each single cell for the validation of gene expression by Real-Time TaqMan single qPCR assays or by TLDA qPCR array.
Zhang Y., Zhong J.F., Qiu H., Robb MacLellan W., Marbán E, & Wang C. (2015). Epigenomic Reprogramming of Adult Cardiomyocyte-Derived Cardiac Progenitor Cells. Scientific Reports, 5, 17686.
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6

Oocyte and Sperm RNA Isolation

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Oocytes and cumulus cells were processed in Cell-to-Ct lysis buffer as recommended (Ambion, #4 458 236, Grand Island, USA) and stored in liquid nitrogen. Discarded sperm samples were pelleted and washed in phosphate-buffered saline (PBS) prior to RNA collection.
Fellmeth J.E., Gordon D., Robins C.E., Scott RT J.r., Treff N.R, & Schindler K. (2015). Expression and characterization of three Aurora kinase C splice variants found in human oocytes. Molecular Human Reproduction, 21(8), 633-644.
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