siRNA targeting MALAT1 (si-MALAT1#1 and si-MALAT1 #2), siRNA negative control (si-NC), pcDNA, and E2F5 were bought from Genepharma (Shanghai, China). miR-1271-5p mimics (miR-1271-5p) and miR-NC, and miR-1271-5p inhibitor (anti-miR-1271-5p) and anti-miR-NC were purchased from RIBOBIO (Guangzhou, China). Cell transfection was performed using Lipofectamine 3000 (Invitrogen).
Si malat1 2
Manufactured by GenePharma
Sourced in China
Si-MALAT1 #2 is a laboratory equipment product from GenePharma. It is designed to facilitate the study of MALAT1, a long non-coding RNA that has been implicated in various cellular processes. The core function of Si-MALAT1 #2 is to enable the silencing or knockdown of MALAT1 expression, which can be used as a research tool to investigate the role of MALAT1 in biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Si malat1 1, by GenePharma (4 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Anti mir nc, by RiboBio (1 mentions)
Negative sirna, by GenePharma (1 mentions)
Mir 1271 5p, by RiboBio (1 mentions)
Anti mir 1271 5p, by RiboBio (1 mentions)
Mir nc, by RiboBio (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Mir 101 3p mimic, by RiboBio (1 mentions)
Mimics nc, by RiboBio (1 mentions)
Lv malat1, by GenePharma (1 mentions)
Sirna against, by GenePharma (1 mentions)
Mir 101 3p inhibitor, by RiboBio (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Si nc, by GenePharma (1 mentions)
Sirna, by GenePharma (1 mentions)
Lv nc, by GenePharma (1 mentions)
Ht 29 atcc htb 38, by American Type Culture Collection (1 mentions)
Hct116 atcc ccl 247, by American Type Culture Collection (1 mentions)
Ccl21, by ZSGB-BIO (1 mentions)
4 protocols using si malat1 2
1
MALAT1 silencing by siRNA
2
Regulatory Mechanisms of MALAT1 in Colon Cancer
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CC cell lines HCT116 (ATCC® CCL-247) and HT29 (ATCC® HTB-38) were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA), and human normal colon epithelial cell line NCM 460 (CM-H203) was purchased from GAINING BIOLOGICAL (Shanghai, China). All cells were cultured in Roswell Park Memorial Institute 1640 Medium (RPMI1640; Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (Gibco, Grand Island, NY, USA) at 37°C with 5% CO2.
The overexpression plasmid of MALAT1 (LV-MALAT1), the small interfering RNA (si-RNA) against MALAT1 (si-MALAT1-1 and si-MALAT1-2), si-RNA against STC1 (si-STC1), and the negative controls (LV-NC and si-NC) were purchased from GenePharma (Shanghai, China). The targeting sequences for si-MALAT1-1, si-MALAT1-2, si-STC1 and si-NC were 5ʹ-GGCAAUGUUUUACACUAUUTT-3ʹ, 5ʹ-CACAGGGAAAGCGAGTGGTTGGTAA-3ʹ, 5ʹ-CTGCTTAAACAAAGCAGTATA-3ʹ and 5ʹ-UUCUCCGAACGUGUCACGUTT-3ʹ, respectively. In addition, miR-101-3p mimics, miR-101-3p inhibitor and the negative controls (mimics NC and inhibitor NC) were purchased from Ribobio (Guangzhou, China). When reaching 80% confluence, cells were transfected with the above agents using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) for 48 h.
The overexpression plasmid of MALAT1 (LV-MALAT1), the small interfering RNA (si-RNA) against MALAT1 (si-MALAT1-1 and si-MALAT1-2), si-RNA against STC1 (si-STC1), and the negative controls (LV-NC and si-NC) were purchased from GenePharma (Shanghai, China). The targeting sequences for si-MALAT1-1, si-MALAT1-2, si-STC1 and si-NC were 5ʹ-GGCAAUGUUUUACACUAUUTT-3ʹ, 5ʹ-CACAGGGAAAGCGAGTGGTTGGTAA-3ʹ, 5ʹ-CTGCTTAAACAAAGCAGTATA-3ʹ and 5ʹ-UUCUCCGAACGUGUCACGUTT-3ʹ, respectively. In addition, miR-101-3p mimics, miR-101-3p inhibitor and the negative controls (mimics NC and inhibitor NC) were purchased from Ribobio (Guangzhou, China). When reaching 80% confluence, cells were transfected with the above agents using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) for 48 h.
3
GC Cell Line Transfection and Maintenance
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Human GC cell line SGC-7901 (American Type Culture Collection, ATCC, Manassas, VA, USA) was maintained in the Roswell Park Memorial Institute (RPMI)-1640 (South Logan, UT, USA) with 10% fetal bovine serum (Tianhang Biotechnology Co., Ltd., Zhejiang, China) and 100 units/ml penicillin/streptomycin in an incubator with 5% CO2 at 37 °C. The medium was renewed every 24—48 h. The cells exhibiting logarithmic growth were subsequently detached using 0.25% trypsin and transfected with CCL21 (150 μg·L−1 CCL21, ZSGB-Bio, Beijing, China), oe-MALAT1-1, si-MALAT1-1, si-MALAT1-2, or si-SRSF1 (the above plasmids designed, synthesized and constructed by Shanghai GenePharma Co. Ltd., Shanghai, China) in accordance with the instructions of the LipofectamineTM2000 (Invitrogen, Carlsbad, CA, USA).
4
MALAT1-miR-26a Interaction in HLECs
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Small interfering RNAs (siRNAs) targeting MALAT1, including siMALAT1-1and siMALAT1-2, were obtained from GenePharma (Shanghai, China). Primary HLECs were transfected with 100 nM MALAT1 siRNAs or negative control siRNA [si-control] individually for 24 h. The siRNA sequences are shown in Table 1 .
In addition, miR-26a mimics, anti-miR-26a, miR-26a mimics negative control, and anti-miR-26a negative control were obtained from GenePharma. The primary HLECs were transiently transfected with 100 nM miR-26a mimics or anti-miR-26a or negative control for 6 h using GenePORTER transfection reagent (GTS, Inc., San Diego, CA, USA).
The MALAT1 sequence was subcloned into the HindIII and EcoRI sites of pcDNA3.1 (Invitrogen, Carlsbad, CA, USA) vector, named pcDNA3.1-MALAT1. The MALAT1 sequence binding miRNA-26a response elements were mutated in which 5'-CUUGUUAUUUUUUACUUGA-3' changed to 5'-ACCACCCCCCCCCCACCAC-3'. The pcDNA3.1-MALAT1 with mutations was named pcDNA3.1-MALAT1-mut (miRNA-26a). The primary HLECs were transfected with pcDNA3.1-MALAT1 and pcDNA3.1-MALAT1-mut in order to achieve the ectopic expression of MALAT1. The primary HLECs were transfected with an empty pcDNA3.1 vector used as a control.
In addition, miR-26a mimics, anti-miR-26a, miR-26a mimics negative control, and anti-miR-26a negative control were obtained from GenePharma. The primary HLECs were transiently transfected with 100 nM miR-26a mimics or anti-miR-26a or negative control for 6 h using GenePORTER transfection reagent (GTS, Inc., San Diego, CA, USA).
The MALAT1 sequence was subcloned into the HindIII and EcoRI sites of pcDNA3.1 (Invitrogen, Carlsbad, CA, USA) vector, named pcDNA3.1-MALAT1. The MALAT1 sequence binding miRNA-26a response elements were mutated in which 5'-CUUGUUAUUUUUUACUUGA-3' changed to 5'-ACCACCCCCCCCCCACCAC-3'. The pcDNA3.1-MALAT1 with mutations was named pcDNA3.1-MALAT1-mut (miRNA-26a). The primary HLECs were transfected with pcDNA3.1-MALAT1 and pcDNA3.1-MALAT1-mut in order to achieve the ectopic expression of MALAT1. The primary HLECs were transfected with an empty pcDNA3.1 vector used as a control.
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