2.6 Real-Time
Quantitative PCR
of MicroRNAs
1. RT primer and miRNA quantification assay for human
miR-221-3p (hsa-miR-221-3p, ID: 00524), miR-155-5p
(ID: 002623), miR-210-3p (ID: 000512), and miR-96-5p
(ID: 000186) (Thermo Fisher Scientific).
2. Human U6 snRNA (ID: 001973) and hsa-let-7c-5p (ID:
000379): used as references to normalize samples amount for
miRNA quantification.
3. TaqMan MicroRNA Reverse Transcription Kit (Thermo Fisher
Scientific).
4. CFX96 Touch Real-time PCR Detection System (Bio-Rad) to
quantify miRNAs content.
5. DNA polymerase enzyme in TaqMan Universal PCR Master
Mix, no AmpErase UNG 2Â (Thermo Fisher Scientific).
2.7 Viability and Cell
Growth
1. Muse Count and Viability Kit (Luminex Corporation, Austin,
Texas, USA) to evaluate effects of argininocalix[4]arene 1 on
U251 cells viability.
2. Z2 Coulter Counter (Coulter Electronics) to monitor cell
growth.
Fig. 1 Possible targeting of anti-apoptotic PUMA mRNA by miR-221-3p (a) and interaction between PNA-a221
and miR-221-3p (b). The PNA sequences discussed in this chapter are shown in Table 1. Other apoptosisassociated mRNAs described to be regulated by miR-221-3p are shown in panel c [65–72]. Modified from
Brognara et al. [46]. The criteria for the design of anti-miRNA PNA molecules are discussed in Note 1
128
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Quantitative PCR
of MicroRNAs
1. RT primer and miRNA quantification assay for human
miR-221-3p (hsa-miR-221-3p, ID: 00524), miR-155-5p
(ID: 002623), miR-210-3p (ID: 000512), and miR-96-5p
(ID: 000186) (Thermo Fisher Scientific).
2. Human U6 snRNA (ID: 001973) and hsa-let-7c-5p (ID:
000379): used as references to normalize samples amount for
miRNA quantification.
3. TaqMan MicroRNA Reverse Transcription Kit (Thermo Fisher
Scientific).
4. CFX96 Touch Real-time PCR Detection System (Bio-Rad) to
quantify miRNAs content.
5. DNA polymerase enzyme in TaqMan Universal PCR Master
Mix, no AmpErase UNG 2Â (Thermo Fisher Scientific).
2.7 Viability and Cell
Growth
1. Muse Count and Viability Kit (Luminex Corporation, Austin,
Texas, USA) to evaluate effects of argininocalix[4]arene 1 on
U251 cells viability.
2. Z2 Coulter Counter (Coulter Electronics) to monitor cell
growth.
Fig. 1 Possible targeting of anti-apoptotic PUMA mRNA by miR-221-3p (a) and interaction between PNA-a221
and miR-221-3p (b). The PNA sequences discussed in this chapter are shown in Table 1. Other apoptosisassociated mRNAs described to be regulated by miR-221-3p are shown in panel c [65–72]. Modified from
Brognara et al. [46]. The criteria for the design of anti-miRNA PNA molecules are discussed in Note 1
128
Alessia Finotti et al.