Synthetic miR-27a mimic (Syn-hsa-miR-27a), miR-27a inhibitor (anti-hsa-miR-27a) or the appropriate scrambled controls (AllStar or mirScript Inhibitor-Negative Control) were purchased from Qiagen (Hilden, Germany). The miR-27a-antisense (MZIP27a-PA-1), the pre-miR-27a expression constructs (PMIRH27a-onlyPA-1) and scrambled control miRNAs (MZIP000-PA-1; PMIRH000PA-1) plasmids (System Biosciences, Mountain View, CA, USA) were transfected in the different CRC cell lines. microRNA functional studies were performed by inhibiting miRNA-mRNA target interactions either with a custom-designed calreticulin-miScript Target Protector or a negative control miScript Target Protector (MTP0075035; Qiagen). Detection of no variations in unrelated proteins validated target protector specificity. A gene-specific package of three preselected siRNAs against calreticulin (Flexi Tube siRNA GS811) or a negative control siRNA (SI03650325) (Qiagen) was used in transient transfections. Functional assays, RNA and protein analyses were performed within 24/72 h from transfections. In each experiment, the extent of miR-27a silencing/overexpression was assessed by qRT-PCR. Plasmids and oligonucleotides were transfected using Lipofectamine 2000 (Thermo Fisher, Waltham, MA, USA) and HiPerFect Transfection Reagent (Qiagen) or RNAi Max (Thermo Fisher), respectively, according to the manufacturers' recommendations.
Si03650325
Manufactured by Qiagen
Sourced in France
The SI03650325 is a laboratory equipment product manufactured by Qiagen. It is designed to perform a specific function, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation or interpretation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Mzip000 pa 1, by System Biosciences (3 mentions)
Medium 199, by Thermo Fisher Scientific (2 mentions)
Interferin, by Polyplus Transfection (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Medium 199, by Merck Group (2 mentions)
Rnaimax, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Miscript target protector, by Qiagen (1 mentions)
Mzip27a pa 1, by System Biosciences (1 mentions)
Pmirh000 pa 1, by System Biosciences (1 mentions)
Hiperfect transfection reagent, by Qiagen (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
4 protocols using si03650325
1
Modulating miR-27a in Colorectal Cancer
2
Rat CD9+ Cells Transfection Protocol
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CD9-positive cells prepared from Wistar rats were plated at 2.0 × 104 cells/cm2 or 1.0 × 105 cells/cm2 on 8-well glass chamber slides with laminin-coated surfaces. Cells were then cultured for 24 h in 400 μL Medium 199 with 10% FBS (Merck Millipore) at 37 °C in a humidified atmosphere of 5% CO2 and 95% air. For transfection of siRNAs, the culture medium was replaced with 400 μL of Medium 199 with 10% FBS (Thermo Fisher Scientific) supplemented with transfection reagent (INTERFERin at 1:100 v/v; PolyPlus Transfection, Illkirch, France) and siRNAs against Cd9 mRNA (0.2 μM Rn_Cd9_1; Qiagen, Venlo, Netherlands) in wells with cells plated at a density of 1.0 × 105 cells/cm2 or siRNAs against Id2 mRNA (0.2 μM Rn_Id2_1; Qiagen) in wells with cells plated at a density of 2.0 × 104 cells/cm2, followed by further cultivation for 48 and 120 h, respectively. A non-silencing siRNA without homology to any known mammalian gene was used as a negative control (SI03650325;Qiagen). After cultivation, cells were used for proliferation assays and immunocytochemistry.
3
Downregulation of RECQ1 and FEN-1 in HeLa Cells
4
Silencing CD9 and CD81 in Mammary Cells
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CD9-positive cells isolated from rat mammary glands were plated at a density of 1.0 × 105 cells/cm2 on 8-well glass chamber slides coated
with laminin. The cells were then cultured at 37°C for 24 h in 400 μl of Medium 199 containing 10% FBS (Merck Millipore) in a humidified atmosphere of 5%
CO2 and 95% air. For small INTERFERing RNA (siRNA) transfection, the culture medium was replaced with 400 μl of Medium 199 containing 10% FBS
(Thermo Fisher Scientific) supplemented with the transfection reagent INTERFERin (1:100 v/v; PolyPlus Transfection, Illkirch, France) and siRNAs against
Cd9 or/and Cd81 (0.2 μM Mm_Cd9_1 and Mm_Cd81_2; Qiagen), and the cells were incubated for
48 h. An siRNA without homology to any known mammalian gene was used as the negative control (SI03650325; Qiagen). The cultivated cells were used for
proliferation assays and immunocytochemistry.
with laminin. The cells were then cultured at 37°C for 24 h in 400 μl of Medium 199 containing 10% FBS (Merck Millipore) in a humidified atmosphere of 5%
CO2 and 95% air. For small INTERFERing RNA (siRNA) transfection, the culture medium was replaced with 400 μl of Medium 199 containing 10% FBS
(Thermo Fisher Scientific) supplemented with the transfection reagent INTERFERin (1:100 v/v; PolyPlus Transfection, Illkirch, France) and siRNAs against
Cd9 or/and Cd81 (0.2 μM Mm_Cd9_1 and Mm_Cd81_2; Qiagen), and the cells were incubated for
48 h. An siRNA without homology to any known mammalian gene was used as the negative control (SI03650325; Qiagen). The cultivated cells were used for
proliferation assays and immunocytochemistry.
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