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Lentivirus - 5. page

qRT-PCR for Lentivirus Titration

1. Make two primer probe sets U5-Psi: U5-Psi forward: TGTGTGCCCGTCTGTTGTGT U5-Psi reverse: GAGTCCTGCGTCGAGAGAGC U5-Psi probe: FAM-CAGTGGCGCCCGAACAGGGA-TAMARA WPRE: WPRE forward: CCGTTGTCAGGCAACGTG WPRE reverse: AGCTGACAGGTGGTGGCAAT WPRE probe: FAMTGCTGACGCAACCCCCACTGGT-TAMARA 2. Purify RNA from […]

Concentrate Lentivirus by Ultra-centrifugation

Ultracentrifuge: Beckman LE 80K Rotor: SW28 Tubes: #326823 autoclaved (polyallomer) 1. Pool all viral media into and filter through a 0.45μm HV Durapore membrane (Millipore #SCHVU01RE). 2. Place tubes into […]

Lentivirus Purification by Ultra-centrifugation

Before Starting: Required equipments and consumbles: 1. SW-28 rotor with buckets and caps (Kept in 4C Refrigerator in Centrifuge Room) 2. SW-28 thin walled ultra clear centrifuge tubes (Beckman # […]

Lentivirus Titration via qRT-PCR

The transduction unit (TU or IFU) of the lentiviral particles was estimated using the formula which 1TU=100 copies of viral genomic RNA. The physical copy numbers of the viral genomic […]