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Dntps

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DNTPs (Deoxynucleotide Triphosphates) are essential building blocks for DNA synthesis. They provide the necessary nucleotides for the polymerization of DNA during various molecular biology and genetic engineering applications.

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Lab products found in correlation

Cytochalasin d, by Merck Group (3 mentions) Jph203, by Selleck Chemicals (3 mentions) Nct 503, by Merck Group (3 mentions) Latrunculin a, by Cayman Chemical (3 mentions) S 5 adenosyl l methionine iodide sam, by Merck Group (3 mentions) Pcw57 gfp p2a mcs, by Addgene (3 mentions) N acetyl l cysteine nac, by Merck Group (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Fibronectin, by Merck Group (3 mentions) Dasatinib, by Selleck Chemicals (3 mentions) Pf 573228, by Selleck Chemicals (3 mentions) 17 aag geldanamycin, by LC Laboratories (3 mentions) Sybr green master mix, by Thermo Fisher Scientific (1 mentions) Rnasin ribonuclease inhibitor, by Promega (1 mentions) Reliaprep rna miniprep system, by Promega (1 mentions) Bigdye 3, by Thermo Fisher Scientific (1 mentions) Biomek 3000, by Beckman Coulter (1 mentions) Mgcl2, by Thermo Fisher Scientific (1 mentions) Amplitaq gold, by Thermo Fisher Scientific (1 mentions)

5 protocols using dntps

1

Ectopic Expression of TP53 Variants

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The following compounds and working concentrations were used: Fibronectin (10 μg/ml, Sigma Aldrich F0895), JPH203 (Selleck Chemicals S8667), NCT503 (Sigma Aldrich SML1659), Dasatinib (Selleck Chemicals S1021), PF-573228 (Selleck Chemicals S2013), Cytochalasin D (Sigma Aldrich C2618), Latrunculin-A (Cayman 10010630), SAHA Cayman 149647-78-9), 17-AAG Geldanamycin (Lc Laboratories A-6880), GCN2-IN-1 (Synonyms: A-92; MedChem Express Cat. No.: HY-100877), nucleosides dNTPs (Euroclone EMR276425), N-Acetyl-L-Cysteine (NAC) (Sigma Aldrich A9165), S-(5′-Adenosyl)-L-Methionine Iodide (SAM) (Sigma Aldrich A4377) and DMSO (Sigma Aldrich D4540). Treatments lasted as described in figure legends.
pCW57-GFP-P2A-MCS (empty backbone) was bought from Addgene (plasmid #89181). pCW57-GFP-HA-p53 wt, pCW57-GFP-HA-mutp53 R175H, pCW57-GFP-HA-mutp53 R249S, pCW57-GFP-HA-mutp53 R273K or pCW57-GFP-HA-mutp53 R280K were generated by cloning a PCR amplified DNA fragment of the human HA-tagged TP53 sequence (WT or mutated) into the pCW57-GFP-P2A-MCS with MluI and BamHI restriction sites.
Tombari C., Zannini A., Bertolio R., Pedretti S., Audano M., Triboli L., Cancila V., Vacca D., Caputo M., Donzelli S., Segatto I., Vodret S., Piazza S., Rustighi A., Mantovani F., Belletti B., Baldassarre G., Blandino G., Tripodo C., Bicciato S., Mitro N, & Del Sal G. (2023). Mutant p53 sustains serine-glycine synthesis and essential amino acids intake promoting breast cancer growth. Nature Communications, 14, 6777.
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2

Ectopic Expression of TP53 Variants

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The following compounds and working concentrations were used: Fibronectin (10 μg/ml, Sigma Aldrich F0895), JPH203 (Selleck Chemicals S8667), NCT503 (Sigma Aldrich SML1659), Dasatinib (Selleck Chemicals S1021), PF-573228 (Selleck Chemicals S2013), Cytochalasin D (Sigma Aldrich C2618), Latrunculin-A (Cayman 10010630), SAHA Cayman 149647-78-9), 17-AAG Geldanamycin (Lc Laboratories A-6880), GCN2-IN-1 (Synonyms: A-92; MedChem Express Cat. No.: HY-100877), nucleosides dNTPs (Euroclone EMR276425), N-Acetyl-L-Cysteine (NAC) (Sigma Aldrich A9165), S-(5′-Adenosyl)-L-Methionine Iodide (SAM) (Sigma Aldrich A4377) and DMSO (Sigma Aldrich D4540). Treatments lasted as described in figure legends.
pCW57-GFP-P2A-MCS (empty backbone) was bought from Addgene (plasmid #89181). pCW57-GFP-HA-p53 wt, pCW57-GFP-HA-mutp53 R175H, pCW57-GFP-HA-mutp53 R249S, pCW57-GFP-HA-mutp53 R273K or pCW57-GFP-HA-mutp53 R280K were generated by cloning a PCR amplified DNA fragment of the human HA-tagged TP53 sequence (WT or mutated) into the pCW57-GFP-P2A-MCS with MluI and BamHI restriction sites.
Tombari C., Zannini A., Bertolio R., Pedretti S., Audano M., Triboli L., Cancila V., Vacca D., Caputo M., Donzelli S., Segatto I., Vodret S., Piazza S., Rustighi A., Mantovani F., Belletti B., Baldassarre G., Blandino G., Tripodo C., Bicciato S., Mitro N, & Del Sal G. (2023). Mutant p53 sustains serine-glycine synthesis and essential amino acids intake promoting breast cancer growth. Nature Communications, 14, 6777.
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3

Ectopic Expression of TP53 Variants

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The following compounds and working concentrations were used: Fibronectin (10 μg/ml, Sigma Aldrich F0895), JPH203 (Selleck Chemicals S8667), NCT503 (Sigma Aldrich SML1659), Dasatinib (Selleck Chemicals S1021), PF-573228 (Selleck Chemicals S2013), Cytochalasin D (Sigma Aldrich C2618), Latrunculin-A (Cayman 10010630), SAHA Cayman 149647-78-9), 17-AAG Geldanamycin (Lc Laboratories A-6880), GCN2-IN-1 (Synonyms: A-92; MedChem Express Cat. No.: HY-100877), nucleosides dNTPs (Euroclone EMR276425), N-Acetyl-L-Cysteine (NAC) (Sigma Aldrich A9165), S-(5′-Adenosyl)-L-Methionine Iodide (SAM) (Sigma Aldrich A4377) and DMSO (Sigma Aldrich D4540). Treatments lasted as described in figure legends.
pCW57-GFP-P2A-MCS (empty backbone) was bought from Addgene (plasmid #89181). pCW57-GFP-HA-p53 wt, pCW57-GFP-HA-mutp53 R175H, pCW57-GFP-HA-mutp53 R249S, pCW57-GFP-HA-mutp53 R273K or pCW57-GFP-HA-mutp53 R280K were generated by cloning a PCR amplified DNA fragment of the human HA-tagged TP53 sequence (WT or mutated) into the pCW57-GFP-P2A-MCS with MluI and BamHI restriction sites.
Tombari C., Zannini A., Bertolio R., Pedretti S., Audano M., Triboli L., Cancila V., Vacca D., Caputo M., Donzelli S., Segatto I., Vodret S., Piazza S., Rustighi A., Mantovani F., Belletti B., Baldassarre G., Blandino G., Tripodo C., Bicciato S., Mitro N, & Del Sal G. (2023). Mutant p53 sustains serine-glycine synthesis and essential amino acids intake promoting breast cancer growth. Nature Communications, 14, 6777.
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4

RNA Extraction, cDNA Synthesis, and qRT-PCR

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Total RNA was purified using ReliaPrep RNA Miniprep System (Promega, #Z6011), according to the manufacturer's instructions. RNA was retrotranscribed for 1 h at 42°C with retro transcriptase M‐MLV (Invitrogen, #28025‐013) in the presence of oligo(dT)15 Primer (Promega, #C1101), dNTPs (Euroclone), and RNAsin Ribonuclease Inhibitor (Promega, #N2111). The cDNA was diluted 10 times to perform qRT‐PCR with SYBR Green Master Mix (Applied Biosystems, #4309155). The list of primers adopted in the study is provided in Appendix Table S2. Ct values were normalized to the housekeeping gene UbiquitinC by the ΔΔCt method.
Perucho L., Icardi L., Di Simone E., Basso V., Agresti A., Vilas Zornoza A., Lozano T., Prosper F., Lasarte J.J, & Mondino A. (2023). The transcriptional regulator Sin3A balances IL‐17A and Foxp3 expression in primary CD4 T cells. EMBO Reports, 24(5), e55326.
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5

Comprehensive Genotyping of CYP3A, ABCB1, and SXR

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As regards genotyping of CYP3A5, ABCB1 and SXR polymorphisms 500 μl of whole blood were collected during routine ambulatory control.
DNA extraction was performed by extractor Fuji QuickGene-810 (Fujifilm, Tokyo, Japan), PCR was carried out in 20 μl of a solution containing 2 μl of 10 x PCR Gold Buffer, 2 mM of MgCl2 (Applied Biosystem, Foster City, CA, USA), 80 μM each of dNTPs (Euroclone, Pero, Milan, Italy), 50 pmol each of primers for CYP3A and ABCB1 as previous described [32 (link)], 50 ng of genomic DNA and 0.6 U of AmpliTaq Gold (Applied Biosystem, Foster City, CA USA). For the polymorphism of SXR A7635G and SXR –200 GAGAAG/− (rs3842689) we used the following primers: SXR A7635G forward 3′- TGG ATG CCA AGC TCA GTGG − 5′; reverse 3′- CAG CAG CCA TCC CAT AAT CC − 5′; for SXR rs3842689 we used the following primers pair: forward 3′-CTG ATG CTC TCT GGT CCT GC − 5′, reverse 3′-TGC CTG CTA TAG CTG ATT CAT TG-5′ with a melt temperature of 60 °C for both polymorphisms..
The template was purified by liquid handling Biomek® 3000 (Beckman Coulter, CA, USA) using a magnetic particles system (Agencourt/Beckman Coulter, CA, USA). The single DNA strand was amplified by BigDye® 3.1 (Applied Biosystems, Foster City, CA USA) and then sequenced by a 3130xl Genetic Analyzer (Applied Biosystems/Hitachi, Foster City, CA USA).
Turolo S., Edefonti A., Ghio L., Testa S., Morello W, & Montini G. (2020). CYP and SXR gene polymorphisms influence in opposite ways acute rejection rate in pediatric patients with renal transplant. BMC Pediatrics, 20, 246.
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