Viral Gene Delivery Technology

pSLIC-PuroP2ATM

AbDia Technologies Inc. provides pSLIC-PuroP2ATM lentivirus plasmid construction kit (LT2002) to let user creates the lentivirus plasmid for the gene of interest. This kit comes with SLIC-EnzymeTM(SE1001-01) and a lentivirus plasmid which contains puromycin resistance gene and P2A sequence.


SLIC (sequence and ligation independent cloning) allows users to insert PCR product into the plasmid without ligation. P2A peptide sequence allows to express both puromycin resistance gene and user’s gene of interest by a single CMV promoter (bicistronic). P2A sequence in our plasmid is GSGATNFSLLKQAGDVEENPG/PD. The first part (before slash "/ ") GSGATNFSLLKQAGDVEENPG is attached to puromycin resistance gene product and the second part (after slash "/ ") PD is attached to the gene of interest users insert. 


Our proprietary SLIC-EnzymeTM system which has superb performance and temperature stability (stable even at room temperature for a few days). In 30min reaction with SLIC-EnzymeTM, PCR product is inserted downstream of P2A sequence.

Components and Storage

Catalogue # Component Size Storage
LT2002-01 pSLIC-PuroP2ATM Plasmid 20µl (200ng/µl) 4°C.This plasmid is not intended to amplify.
SE1001-01 10×SLIC-EnzymeTM 25µl -20°C, Enzyme is stable at least 4 days at room temperature.
GFP1001-01 GFP Template DNA (control) 10µl (10ng/µl) 4°C
GFP1002-01 GFP Forward and Reverse Primer Mixture 10µl, 10µM 4°C 

The kit contains enough plasmids and reagents to perform at least 20 reactions for creating lentiviral vector plasmids which contain gene of interest.

Kit Components and Storage Conditions

Catalogue # Component Size Storage Condition
LT1001-1 Lentivirus production medium 55mL

4 °C. Add 1.2ml FBS before use.
LT1001-2 Helper-plasmids mixture 0.2mL 4 °C
LT1001-3 Production helper-solution 0.5mL 4 °C
LT1001-4 Plate coating solution 2.0mL 4 °C
LT1001-5 Lentivirus Purification Solution 20mL 4 °C
LT1001-6 Lentivirus storage solution 2.0mL 4 °C
LT1001-7 GFP expressing lentivirus plasmid 1µg in 20µl TE 4 °C. Ampicillin resistance

References

  1. Y. Ido, et al. PLoS ONE, 2012 Apr 07(4): e35092

  2. Mamie Z Li & Stephen J Elledge Nature Methods 2007; 4:251-256

  3. Gibson DG Methods Enzymol 2011; 498:349-61

  4. Jin Hee Kim, Sang-Rok Lee, Li-Hua Li, et al. PLoS ONE 2011 Apr 06 (4): e18556

  5. Georgios Trichas, Jo Begbie and Shankar Srinivas BMC Biology 2008, 6:40