Total RNA was extracted using Trizol reagent (Invitrogen) following the protocol. The SYBR green PCR Master Mix (Qiagen, Hilden, Germany) was used for mRNA expression detection following the protocol using GAPDH expression as an endogenous control. The expression of miRNA was examined by a Hairpinit™ miRNAs qPCR kit (Genepharma, Shanghai, China) using RNU6B as an endogenous control. The 2−ΔΔCT method was used to analyze the relative fold changes.
Hairpin it mirnas qpcr kit
Manufactured by GenePharma
Sourced in China
The Hairpin-it TM miRNAs qPCR kit is a laboratory equipment product designed for the quantitative analysis of microRNA (miRNA) expression using real-time PCR (qPCR) technology. The kit provides the necessary reagents and primers to detect and quantify specific miRNA targets in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (27 mentions)
Sybr green qpcr assay, by Takara Bio (12 mentions)
Sybr green pcr master mix, by Qiagen (8 mentions)
Sybr green supermix, by GenePharma (2 mentions)
Dnase 1, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
One step sybr primerscript rt pcr kit, by Takara Bio (1 mentions)
Abi prism 7900ht, by Thermo Fisher Scientific (1 mentions)
Access rt pcr kit, by Promega (1 mentions)
Sybr green quantitative rt qpcr kit, by Merck Group (1 mentions)
Direct zol rna kit, by Zymo Research (1 mentions)
Dnase 1, by Merck Group (1 mentions)
Sybr green qrt pcr assay, by Takara Bio (1 mentions)
Riboex, by Thermo Fisher Scientific (1 mentions)
Superscript 2, by Thermo Fisher Scientific (1 mentions)
Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Reverse transcript kit, by Takara Bio (1 mentions)
Sybr green realtime pcr kit, by Qiagen (1 mentions)
30 protocols using hairpin it mirnas qpcr kit
1
Comprehensive mRNA and miRNA Expression Analysis
2
Quantification of miRNA and mRNA Expression in Colorectal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from CRC cells using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). The expression levels of miRNAs were detected using the Hairpin-it miRNAs qPCR Kit (Genepharma, Shanghai, China). The expression of RNU6B served as an endogenous control. iASPP expression was measured using a SYBR Green qPCR assay (Takara, Dalian, China). The data were processed according to the 2−ΔΔCT method; the relative expression of miR-124 was calculated with the formula, 2−(CTmiRNA−CTRNU6B). The primers are shown in Supplementary Table 1 .
3
RNA Extraction and miRNA/mRNA Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from CRC cells using TRIzol reagent (Invitrogen, CA, USA). Total RNA was extracted from tissue samples following the protocol by Helen Pearson [23 (link)]. The expression levels of miR-124 and miR-155 were detected using the Hairpin-it™ miRNAs qPCR Kit (Genepharma, Shanghai, China). The expression of RNU6B served as an endogenous control. IASPP, p63 and STAT1 expression was measured using a SYBR Green qPCR assay (Takara, Dalian, China). The data were processed according to the 2−ΔΔCT method. The primers were shown in Supplementary Table 1 .
4
RNA Extraction and Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA extraction was conducted by employing TRIzol™ (Invitrogen), following the methods described before [27 (link)] with SYBR Green PCR Master Mix (Qiagen) for mRNA expression level determination and a Hairpin-it miRNAs qPCR kit (GenePharma) for miRNA expression level determination. The expression levels of GAPDH and RNU6B were used as endogenous controls. The primer sequences for PCR were shown in Table 1 . Finally, the data were processed using the 2−ΔΔCt relative expression method. The more method details of real time PCR assay listed in Additional file 1 .
5
Quantitative Analysis of miRNA and Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total RNA was extracted from cells using TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc.) and mature miR-185 expression was detected in cells using a Hairpin-it™ miRNAs qPCR kit (Shanghai Genepharma Co., Ltd.). Thermocycling conditions included an initial step at 98°C for 1 min, and 40 cycles at 95°C for 5 sec and at 62–68°C for 15 sec, 72°C for 2 min and final extension at 72°C for 10 min. The expression of small nucleolar RNU6B was used as an endogenous control. SOX9, β-catenin and c-Myc expression was measured using a SYBR green qPCR assay (One Step SYBR PrimerScript RT-PCR Kit, Takara Biotechnology Co., Ltd., Dalian, China). Reverse transcription and amplification were performed at 42°C for 15 min, followed by 40 cycles of 95°C for 5 sec and 60°C for 34 sec using an ABI Prism 7900 HT (Applied Biosystems; Thermo Fisher Scientific, Inc.). The following primer sequences were used for the PCR: RNU6B, reverse 5′-AACGCTTCACGAATTTGCGT-3, forward 5′-CTCGCTTCGGCAGCACA-3; SOX9, forward 5′-AGCGAACGCACATCAAGAC-3′, reverse 5′-CTGTAGGCGATCTGTTGGGG-3′; β-catenin, forward 5′-AAAGCGGCTGTTAGTCACTGG-3′, reverse 5′-CGAGTCATTGCATACTGTCCAT-3′; c-Myc, forward 5′-GGCTCCTGGCAAAAGGTCA-3′, reverse 5′-CTGCGTAGTTGTGCTGATGT-3′; GAPDH, forward 5′-GGAGCGAGATCCCTCCAAAAT-3′, reverse 5′-GGCTGTTGTCATACTTCTCATGG-3′. Data were analysed using the 2−ΔΔCq method (21 (link)).
6
Quantifying miR-133b and Sirt1 mRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from cells using TRIzol reagent (Invitrogen). Mature miR-133b expression in cells was detected using a Hairpin-it™ miRNAs qPCR Kit (GenePharma). Expression of RNU6B was used as an endogenous control. Sirt1 mRNA expression was measured by SYBR Green qPCR assay (Takara, Dalian, P.R. China). Data were processed using the 2−ΔΔCT method.
7
Quantification of MACC1 and miR-574-5p in Colorectal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from colorectal cancer cell lines (SW1116 and HCT116) with TRIzol reagent (Invitrogen, CA, USA). Housekeeping gene of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as internal control. MACC-1 mRNA level was measured with Access RT-PCR kit (Promega, USA). Hairpin-it miRNAs qPCR kit (GenePharma, China) was used to detect the hsa-miR-574-5p expression in SW1116 and HCT116 cell lines with stem-loop reverse transcription. The primers used in the study were shown as follows: MACC-1 (3′UTR sequence) primers, forward 5′-TTGCGGAGGTCACCATAGC-3′; reverse 5′-TTTCCAACAACGGGCTCA-3′; GAPDH primers, forward 5′-GGGCTGCTTTTAACTCTG-3′; reverse 5′-TGGCAGGTTTTTCTAGACGG-3′.
8
Quantifying miR-214 and LINC0086 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from cells using TRIzol reagent (Invitrogen). miR-214 expression in cells was detected using a Hairpin-it™ miRNAs qPCR kit (GenePharma) according to the manufacturer’s instructions. Expression of RNU6B was used as an endogenous control. The expression of LINC0086 was measured by SYBR green qPCR assay (Takara, Dalian, P.R. China) according to the manufacturer’s instructions. Expression of β-actin was used as an endogenous control. qPCR was performed at the following conditions: 95°C for 3 min, and 39 cycles of 95°C for 10 s and 60°C for 30 s. Data were processed using the 2−ΔΔCT method.
9
Quantifying miR-320a and SP1 in CRC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was isolated from CRC tissues and cells using TRIzol® reagent (Invitrogen, Waltham, US). Mature miR-320a expression in cells was determined using a Hairpin-it™ miRNAs qPCR kit (Genepharma, Shanghai, China). RNU6B was used as an endogenous control. The primer sequence for miR-320a was as follows: Forward: 5′-ATGAGAAAAAGCTGGGTTGAGA-3′, reverse: 5′-TATGGTTTTGACGACTGTGTGAT-3′. The primer sequence for RNU6B was: Forward: 5′-CAGCACATATACTAAAATTGGAACG-3′, reverse: 5′-ACGAATTTGCGTGTCATCC-3′. SP1 mRNA expression was determined using the SYBR green qPCR assay (Takara, Dalian, China). GAPDH was used as an endogenous control. The thermocycling conditions were as follows: Denaturation at 95 ˚C for 3 min followed by 40 cycles of amplification at 95 ˚C for 12 s and extension at 62 ˚C for 40 s. GAPDH was used as the endogenous control. Data were analyzed using the 2−ΔΔCt method. The primer sequence for SP1 was as follows: Forward: 5′-TCATACTGTGGGAAACGCTT-3′, reverse: 5′-GACACTCAGGGCAGGCAAA-3′. The primer sequence for GAPDH was: Forward: 5′-TGACTTCAACAGCGACACCCA-3′, reverse: 5′-CACCCTGTTGCTGTAGCCAAA-3′.
10
Quantifying miR-320a and SP1 in CRC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was isolated from CRC tissues and cells using the TRIzol® reagent (Invitrogen, Waltham, US). Mature miR320a expression in cells was determined using a Hairpin-it™ miRNAs qPCR kit (Genepharma, Shanghai, China). RNU6B was used as an endogenous control. SP1 mRNA expression was determined by using SYBR green qPCR assay (Takara, Dalian, China). The thermocycling conditions were as follows: Denaturation at 95˚C for 3 min, followed by 40 cycles of ampli cation at 95˚C for 12 sec and extension at 62˚C for 40 sec. GAPDH was used as the endogenous control. Data was analyzed using the 2 ΔΔCt method.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!