Total RNA was extracted, treated with DNaseI and reverse transcribed using a RNA isolation and cDNA synthesis kit as recommended by the manufacturer (Quiagen, Hilden, Germany). Aliquots of cDNA were employed for Fast SYBR green qPCR (Applied Biosystems, Foster City, USA) 2 and quantified with the 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, USA). The following probes, forward and reverse primer sequences were used: CD32b: 5’- CCATCTGGACTGGAGCCAAC-3’ and 5’- TGGCTTGCTTTTCCCAATGC-3’; CD146: 5’-CGGGTGTGCCAGGAGAG-3’ and 5’-ACCAGTCCACTTGGCTGAAG-3’; vWF: 5’-TGGCAAGGTTTTTCAGGGGA-3’ and 5’-TAAGCAGGTGATGCAGAGGC-3’; Stab-1: 5’-ACGCTTCTAACGCCACCTTT-3’ and 5’-CCACACGATGACGTGGCTAA-3’; Stab-2: 5’-CACTATGTCGGGGATGGACG-3’ and 5’-GGGAGCGTAGGTGGAATACG-3’; LYVE-1: 5’-TACAGGACCCATGGCTGAGA-3’ and 5’-GGTGCCAAGCATTTCGGTTT-3’; RhoA: 5’-CCATCATCCTGGTTGGGAAT-3’ and 5’-CCATGTACCCAAAAGCGC-3’.
Rna isolation and cdna synthesis kit
Manufactured by Qiagen
Sourced in Germany
The RNA isolation and cDNA synthesis kit is a laboratory product designed to extract and convert RNA molecules into complementary DNA (cDNA) for further analysis or applications. This kit provides the necessary reagents and protocols to perform these fundamental molecular biology procedures.
Automatically generated - may contain errors
2 protocols using rna isolation and cdna synthesis kit
1
Quantitative gene expression analysis
2
Axl Gene Expression Quantification
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Axl primers (forward: 5'-CCGGCTGGC-GTATCAAGGCC -3'; reverse: 5'-TGGCTGT-GCTTGCCCTGGG -3'). RNA was extracted, treated with DNaseI and reverse transcribed using a RNA isolation and cDNA synthesis kit (Quia- gen, Hilden, Germany) as recommended by the manufacturer. A SYBR ® Premix Ex Taq ™ ii (Perfect Real Time) Kit (Takara Bio, Shiga, Japan) was used to quantify the Axl mRNA and the GAPDH mRNA via the PCR amplification, as previously described.
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