building 3

Projects

Project: vectors for knocking down gene of interest using shRNA (expressing from U6/H1 polIII promoters- PRC1 and PRC2 Sigma-Aldrich libraries)

ConsultationNo charge
Virus design, per project$200
Amplification of the clones (Midi Preps)$200/clone

Generating two validated positive clones and one validated negative (out-of-target) control clone at the following amounts:

  • 1x107 vg/mL 10mL- Small prep (good for in vitro study):                
  • USC-customers $200/clone ; External Academic-$250/clone including p24 ELISA titering
  • 1x109 vg/mL 20uL- Midi prep (good for two to three in vivo brain injections):
  • USC-customers $600/clone, External Academic-$720/clone including p24 ELISA titering and RCR-safety tests)
  • 1x109 vg/mL 100uL- Large prep (good for 10 to 15 in vivo brain injections):
  • USC-customers $1750/clone, External Academic-$2100/clone including p24 ELISA titering, real-time PCR titering and RCR-safety tests)

Project will be completed in 4-6 weeks for concentrating (medium and large) preps

In vitro validation of the knockdown is $150/clone (Western blotting) (price excludes cost of the antibodies).

Project’s price excludes purchasing shRNA clones from the Sigma-Aldrich (cost is $125/validated clone)


Project: vectors for knocking down gene of interest using miR RNA (expressing from CMV polII promoter- GIPZ TermoFisher libraries or TRE (tet-inducible promoter)

Please keep in mind that clones of this library are not validated, so it is strongly recommended to validate clones (GFP expression and Western blotting) before proceeding to in vivo experiments.

ConsultationNo charge
Virus design, per project $200
Amplification of the clones (Midi Preps)  $200/clone

Generating three-to six clones (will be decided accordingly with the request) available for the fix price of $432/set from the TermoFisher (I will charge $450/set to absorb cost of the out-of-target control which is not provided with the clones) at the following amounts:

  • USC-customers $200/clone; External Academic-$250/clone including p24 ELISA titering

  • 1x109 vg/mL 20uL- Midi prep (good for two to three in vivo brain injections):
  • USC-customers $600/clone, External Academic-$720/clone including p24 ELISA titering and RCR-safety tests)

  • 1x109 vg/mL 100uL- Large prep (good for 10 to 15 in vivo brain injections):
  • USC-customers $1750/clone, External Academic-$2100/clone including p24 ELISA titering, real-time PCR titering and RCR-safety tests)

Project will be completed in 4-6 weeks for concentrating (medium and large) preps

In vitro validation of the knockdown is $150/clone (GFP expression; Western blotting) (price excludes cost of the antibodies)-optional as in/out the core, but recommended.

Project’s price excludes purchasing shRNA clones from the TermoFisher (cost is $450/set of three to six).


Generating miR-employed knockdown with Invitrogen Gateway System Knockdown lentivirus package

  • Includes design, cloning of target gene cDNA into screening plasmid,
  • purchase of miR or shRNA oligonucleotides,
  • cloning of oligonucleotides into appropriate plasmids,
  • screening assay,
  • production of one knockdown virus and a control virus;
  • does not include purchase of target gene cDNA $5000
  • Cloning of miR or shRNA candidate sequences
  • Simultaneous cloning of 4 candidate sequences into miR or shRNA vectors for screening;
    • price does not include purchase of oligonucleotides $1250
    • Screening of miR or shRNA candidate sequences
    • Testing activity of cloned candidate sequences against lacZ-target cDNA;
    • price does not include cloning of target cDNA $1000

     Project, part I: Cloning of the gene of interest cDNA into lentiviral vector, HSV or AAV vectors

    • Cloning design, per project $200
    • Single-step cloning—first plasmid
      Subcloning of cDNA into lentivirus transfer vector AAV or HSV shuttle plasmid $850
    • Single-step cloning—additional plasmid $600
    • Cloning involved multiple steps, cloning large fragments (more than 10 kb), PCR amplification of the insert, or genetic mutagenesis $1250
    • Plasmid prep
    • For plasmids provided by another lab $200
    • Resulted clones will be confirmed by restriction enzyme digestion and sequencing $100

    Project, part IIa:

    • Generating lentiviral vectors
      • 1x107 vg/mL 10mL- Small prep (good for in vitro study): USC-customers $200/clone; External Academic-$250/clone including p24   ELISA titering
      • 1x109 vg/mL 20uL- Midi prep (good for two to three in vivo brain injections): USC-customers $600/clone, External Academic-$720/clone including p24 ELISA titering and RCR-safety tests)
      • 1x109 vg/mL 100uL- Large prep (good for 10 to 15 in vivo brain injections): USC-customers $1750/clone, External Academic-$2100/clone including p24 ELISA titering, real-time PCR titering and RCR-safety tests)

    Project, part IIb: Recombinant HSV1 generation and production

    • Generation and purification of recombinant HSV
    • Transfection of linearized shuttle plasmid and viral DNA; purification through >3 rounds of limiting dilution, expansion culture for subsequent production $3000
    • Concentrated HSV stock, 500 μL of 107 pfu/μL
    • High concentration stock produced from existing expansion culture $1000

    Project, part IIc: Generating Adeno-Associated vectors
    As described in Services and Fees


    Other services

    • p24 ELISA $500/plate, additional plate 250$
    • Competent cells HB101 1ml (20x50ul) >108cfu/μg $150
    • Cesium Grade DNA (to amplify low copy plasmids, DNA for transfection or if required by protocol) 1mg $750