1) Prepare 6 ml of 5X ISO Buffer in a 15 ml falcon tube as follows: 3 ml 1 M Tris-HCl pH 7.5 + 150µl 2 M MgCl2 + 240 µl 100 […]
Monthly Archives: October 2015 - 9. page
Rapid one-step LR recombinational cloning using an enzyme mixture containing Int, IHF and Xis (Gateway Clonase) and two step PCR amplification
Gene specific primers: GSS-F: SbfI Kozak GSS AGGCT cctgcagg CCACCATG NNNNNNNNNNNNNNNNNNNNN GSS-R: GSS * Xho1 (NNNNNNNNNNNNNNNNNNNNN TAG ctcgag ACCCAGCTTTC) For example: GAAAGCTGGGT ctcgag CTA N(15–25) Universal Primers attL1-F ccccGATGAGCAATGCTTTTTTATAATGCCAACTTTGTACAAAAAAGCA […]
Home-made Gateway Clonase II
1.Excision reaction. 44.5 mM Tris–borate (pH 8.3) 7.5 mM Tris–HCl (pH 8.0) 1.15 mM EDTA 5 mM spermidine 60 mM KCl 0.275 mg/ml BSA 15% (v/v) glycerol. Int 50~100 nM […]
SignaGen’s In Vitro siRNA Transfection Reagents
siRNA Transfection Reagent Selection Chart. Reagents Features Transfection Type Toxicity PepMute™ – Exceptional good for siRNA transfection – OK for DNA/siRNA co-transfection siRNA (Yes) DNA/siRNA (No) ++ GenMute™ – Exceptional […]
>300 Pre-made Ready to Use Adenoviruses from SignaGen
SiganGen is offering >300 pre-made and ready to use adenovirus vectors. All the pre-packaged adenoviruses were packaged from Ad.MAX™ system with a serotype 5 adenoviral backbone* (E1/E3 deletion). All the […]
SignaGen’s rAAV Production Service
Multi Approaches to Produce Super Infectious rAAV Particles at Super High Titer: Not like adenovirus, it is relatively harder to achieve high yield of rAAV with regular rAAV production procedures. […]
SignaGen’s In Vitro DNA Transfection Reagents
Transfection Reagent Selection Chart. Reagents Features Transfection Type Toxicity* CalFectin™ – Traditional recipe with new formulation – Most affordable and efficient – Best for large scale virus, antibody and protein […]
One-step qRT-PCR for Lentivirus Titration
1. Extract RNA using a commercial kit 2. DNase treatment using RNase free DNase 1U/ul from Promega to digest residual plasmid DNA 3. Use ABI TaqMan One-Step RT-PCR master mix […]
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 […]