qPCR Lentivirus Titer Kit
The only kit on the market with complete elimination of NTC
abm’s qPCR Lentivirus Titer Kit is a one-step assay which employs a quick RNA extraction step that is followed by RT-qPCR. Designed to deliver high sensitivity and specificity, the kit ensures minimal non-specific background and better overall performance compared to similar kits on the market. ROX reference dye is provided separate from the MasterMix, making this kit universally compatible with most qPCR instruments.
|Unit quantity||100 rxn|
qPCR Lentivirus Titer Kit comes with a separate vial of ROX Reference Dye which can be added depending on the qPCR machine type, as listed in the table below.
Store at -20°C. This product is stable for 2 year from the date of shipping if stored and handled properly.
- qPCR Lentivirus Titer Calculation Form
- Why choose abm's PCR Enzymes?
- qPCR Lentivirus Titer Kit (Flyer)
- PCR Product Guide (2021)
|Why do I get higher titers for the more diluted sample from the same stock?|
This means your virus stock may have strong PCR inhibitors in the sample. Thus, it would best to trust the more diluted samples' results, as it lessens the inhibitor's influence.
|Will the transfected plasmids used to produce the lentiviral particles interfere with the qPCR results? What about the host's genome?|
The transfection plasmids and host's genome remain in the packaging cells while the viral particles are harvested from the supernatant. The titering process will not involve the cells, thus the plasmids and genome will not interfere with the results as well.
|Can this kit titer FIV particles?|
No, this kit cannot titer FIV particles.
|My experiment needs MOI=10. How can I know that viral particle? How to IU(infectious units)/ml convert to viral particle?|
Multiplicity of Infection (MOI) Calculation Method: MOI = (Product Titer (IU/ml) x Virus Volume (ml)) /Total Cell Number For example, to achieve a MOI of 10 with a 1 x 10^6 IU/ml titer virus on 1 x 10^5cells, use 1.0 ml of the virus.
|Does your titration kit work with pLKO.1 based siRNA lentivirus?|
Our titer kit should be able to detect pLKO.1 vector with an qPCR product size of about 100 bp.
|I am interested in finding a kit to titrate lentivirus. I am using a VSVG, RRE, REV packing system with an FUGW construct. I have had trouble in the past with these type of kits since not all lentiviruses contain the same genes. Do you know if your kit works for this specific packaging system?|
According to the sequence of the FUGW plasmid, abm's qPCR Lentivirus Titration Kit is compatible with FUGW plasmid with an amplicon size of around 120bp.
|Is this kit suitable for titrating retroviruses and adenoviruses?|
No, this kit is designed for Lentiviruses only.
|The dissociation curve shows that the Tm of my Lentivirus is lower than that of standards, why?|
The Tm of a qPCR product is mainly depending on two variables: the reaction mixture's chemical compositions and the sequence of the amplicon. Since all the reactions in this case have almost the same compositions (ex: all the input templates are basically water, not for instant high salt template solution), the main variation would be in the template sequence. It is possible that your the vector has a slightly different sequence (likely having less GC content) than our std's sequence, thus having a lower Tm than the Std.
|Will the qPCR Lentivirus Titration(Titer) Kit amplify the integrated genomic region of the lentivirus?|
No, the qPCR Lentivirus Titration (Titer) Kit (Cat#LV900) will not amplify the integrated genomic regions of lentivirus because the primers amplify the 5' LTR of the lentivirus.
|Does the assay detect free viral RNA and so overestimate the viral titer? If so, is there a way to minimize this?|
In theory, it is possible for our Cat#LV900 to detect free viral RNA and free viral RNA can be present in the supernatant collected during lentivirus production, however this should not impact the titer to any significant extent and should be very minimal in detection, if any. If desired, you may wish to further purify your virus using our Ultra-Pure Lentivirus Purification Kit (Cat#LV998) or Speedy Lentivirus Purification (Cat#LV999) to remove any free viral RNA. However, we have never encountered any issues using our LV900 kit and free viral RNA has not been a factor; we use this kit regularly during our routine QC processes, and it is one of the most popular products in our catalogue.
|For lenti titering, is it possible to start with an ultracentrifuged lenti sample (suspended in PBS) as opposed to straight supernatant?|
Yes, starting with an ultracentrifuged lenti sample should indeed be suitable for use with our qPCR Lentivirus Titration(Titer) Kit (LV900).
|Is this kit suitable for detecting both 2nd and 3rd generation lentivirus?|
Yes, the kit is suitable for detecting both 2nd and 3rd generation lentivirus.
|What type of vectors will this kit work with?|
The titer primers supplied in LV900 are designed for all HIV-1 based viral detection.
|Is DNAse treatment required after harvesting the lentivirus stock?|
DNase treatment is not recommended. When packaging lentivirus, we recommend removing the transfection medium from the cells after overnight incubation, and replace with complete culture media for the cells. Incubate another 24 hours, and then collect the supernatant.
|What is the primer design based on?|
The primers in this kit uses a set of proprietary sequences designed based on the 5'LTR region.
|How come my cells are not successfully infected with the evaluated titer from the kit?|
One important thing to note is that the infection success is dependent on the cell type. Some cells may be more susceptible than others for viral infection. Thus, the same number of infectious units may perform differently on different cell types.
|What is the sensitivity of the qPCR Lentivirus Titer Kit (LV900)?|
This kit can detect as low as 10^4 IU/ml.
- Taylor, HE et al. "The innate immune factor apolipoprotein L1 (APOL1) restricts HIV-1 infection" J of Virol. : (2013). PubMed: 24173214.. Application: Lentivirus Titer.
- Thomas, AM et al. "Polysaccharide-modified scaffolds for controlled lentivirus delivery in vitro and after spinal cord injury" J Control Release 170 (3):421-429 (2013). DOI: 10.1016/j.jconrel.2013.06.013. Application: Lentivirus Titer.
- Yu, X et al. "Short-chain fatty acids from periodontal pathogens suppress histone deacetylases, EZH2, and SUV39H1 to promote Kaposi's sarcoma-associated herpesvirus replication" J. Virol. 88 (8):4466-4479 (2014). DOI: 10.1128/JVI.03326-13. PubMed: 24501407. Application: Lentivirus Titer.
- WZang, ZY et al. " Immortalized porcine liver sinusoidal endothelial cells: an in vitro model of xenotransplantation-induced thrombocytopenia" Xenotransplantation 19 (4):249-255 (2012). DOI: 10.1111/j.1399-3089.2012.00715.x. PMID: 22909138. Application: Lentivirus Titer.
- Hbattacherjee, A et al. "Bone morphogenetic protein 4 mediates estrogen-regulated sensory axon plasticity in the adult female reproductive tract" J Neurosci 33 (3):1050-1061a (2013). DOI: 10.1523/JNEUROSCI.1704-12.2013. PubMed: 23325243. Application: Lenti Titer.
- Gower, RM et al. "Modulation of leukocyte infiltration and phenotype in microporous tissue engineering scaffolds via vector induced IL-10 expression" Biomaterials 6 (35):2024-2031 (2014). DOI: 10.1016/j.biomaterials.2013.11.036. PubMed: 24309498. Application: Lentivirus Titer.
- Bhattacherjee, A et al. "Bone morphogenetic protein 4 mediates estrogen-regulated sensory axon plasticity in the adult female reproductive tract" J. Neurosci. 3 (33):1050-1061a (2013). DOI: 10.1523/JNEUROSCI.1704-12.2013. PubMed: 23325243. Application: Lentivirus Titer.
- Azarin, S. M. et al. "In vivo capture and label-free detection of early metastatic cells" Nat. Commun. : (2015). DOI: 10.1038/ncomms9094.
- Chahine, NO et al. "Effect of age and cytoskeletal elements on the indentation-dependent mechanical properties of chondrocytes" PLoS ONE 8 (4):e61651 (2013). DOI: 10.1371/journal.pone.0061651. PubMed: 23613892. Application: Lentivirus Titer.
- Boehler, RM et al. "Lentivirus delivery of IL-10 to promote and sustain macrophage polarization towards an anti-inflammatory phenotype" Biotechnol. Bioeng. 111 (6):1210-1221 (2014). DOI: 10.1002/bit.25175. PubMed: 24375008. Application: Lentivirus Titer.
- Chen, YL et al. "Small interfering RNA targeting nerve growth factor alleviates allergic airway hyperresponsiveness" Mol Ther Nucleic Acids 3:e158 (2014). DOI: 10.1038/mtna.2014.11. PubMed: 24714423. Application: Lentivirus Titer.
- Thomas, AM et al. "Sonic hedgehog and neurotrophin-3 increase oligodendrocyte numbers and myelination after spinal cord injury" Royal society of chemistry : (2014). DOI: 10.1039/C4IB00009A. PubMed: 24873988. Application: Lentivirus Titer.
- George, R et al. "A SHORT INTERFERING RNA MOLECULAR BEACON FOR THE ATTENUATION OF MYCOBACTERIAL INFECTION" American Journal of Biochemistry and Biotechnology 10:40-49 (2014). DOI: 10.3844/ajbbsp.2014.40.49. Application: Viral Titer Determination.
- Yu, X et al. "Short-Chain Fatty Acids from Periodontal Pathogens Suppress Histone Deacetylases, EZH2, and SUV39H1 To Promote Kaposi's Sarcoma-Associated Herpesvirus Replication" J Virol. 88(8):4466-79 (2014). DOI: 10.1128/JVI.03326-13. PubMed: 24501407. Application: Viral Titer.
- Puppo, A et al. "Retinal transduction profiles by high-capacity viral vectors" Gene Ther. 21(10):855-65 (2014). DOI: 10.1038/gt.2014.57. PubMed: 24989814. Application: Lentivirus Titer.
- Chakraborty, S et al. "A CRISPR/Cas9-based system for reprogramming cell lineage specification" Stem Cell Reports 3(6):940-7 (2014). PubMed: 25448066.
- Mak, IW et al. "The epigenetic regulation of SOX9 by miR-145 in human chondrosarcoma" J Cell Biochem 116(1):37-44 (2015). DOI: 10.1002/jcb.24940. PubMed: 25145279.
- Sundararaman, B. et al. "Resources for the comprehensive discovery of functional RNA elements" bioRxiv beta: The Preprint Server for Biology : (2015). DOI: http://dx.doi.org/10.1101/030486.
- Thomas, A. M. et al. "Sponge-mediated lentivirus delivery to acute and chronic spinal cord injuries" J Control Release :1-10 (2015). DOI: 10.1016/j.jconrel.2015.02.032. PubMed: 25724274.
- Sheu, J. et al. "Large-scale production of lentiviral vector in a closed system hollow fiber bioreactor" Mol. Ther. Methods Clin. Dev. : (2015). DOI: doi:10.1038/mtm.2015.20.
- Fransson, M. et al. "Mesenchymal stromal cells support endothelial cell interactions in an intramuscular islet transplantation model" Regen Med Res. 1: (2015). DOI: 10.1186/s40340-015-0010-9.
- Cao, Z. et al. "Ubiquitin Ligase TRIM62 Regulates CARD9-Mediated Anti-fungal Immunity and Intestinal Inflammation" Immunity. 4:715-726 (2015). DOI: doi: 10.1016/j.immuni.2015.10.005. PubMed: 26488816.
- Krassnig, S. et al. "Influence of Lentiviral β-Synuclein Overexpression in the Hippocampus of a Transgenic Mouse Model of Alzheimer's Disease on Amyloid Precursor Protein Metabolism and Pathology" Neurodegener Dis. 4:243-257 (2015). DOI: 10.1159/000430952. PubMed: 26111745.
- Das, B. et al. "Short chain fatty acids potently induce latent HIV-1 in T-cells by activating P-TEFb and multiple histone modifications" Virology. :65-81 (2015). DOI: 10.1016/j.virol.2014.10.033. PubMed: 25463605.
- Shang, R. et al. "Ribozyme-enhanced single-stranded Ago2-processed interfering RNA triggers efficient gene silencing with fewer off-target effects" Nat. Commun. : (2015). DOI: doi:10.1038/ncomms9430.
- Zhang, M et al. "Epigenetically mediated spontaneous reduction of NFAT1 expression causes imbalanced metabolic activities of articular chondrocytes in aged mice" Osteoarthritis and Cartilage 24.7:1274–1283 (2016). DOI: 10.1016/j.joca.2016.02.003. Application: Titer Determination.
- Robitaille, K et al. "High-throughput Functional Genomics Identifies Regulators of Primary Human Beta Cell Proliferation" The Journal of Biological Chemistry 291.9:4614-4625 (2016). DOI: 10.1074/jbc.M115.683912. Application: Titer Determination.
- Theis, et al. "Lentiviral Delivery of miR-133b Improves Functional Recovery After Spinal Cord Injury in Mice" Mol. Neurobiol. : (2016). DOI: 10.1007/s12035-016-0007-z. Application: Titer Determination.
- Henry, A et al. "New role of osteopontin in DNA repair and impact on human glioblastoma radiosensitivity" Oncotarget 2016:1-14 (2016). Application: Titer Determination.
- Margul, D et al. "Reducing neuroinflammation by delivery of IL-10 encoding lentivirus from multiple-channel bridges" Bioengineering and Translational Medicine 1.2:136-148 (2016). DOI: 10.1002/btm2.10018.
- Jin, C et al. "Safe engineering of CAR T cells for adoptive cell therapy of cancer using long-term episomal gene transfer" EMBO Mol. Med. 8.7:702-711 (2016). DOI: 10.15252/emmm.201505869. Application: Production and Titration of Vectors.
- Kurata, M et al. "Using genome-wide CRISPR library screening with library resistant DCK to find new sources of Ara-C drug resistance in AML" Sci. Rep. 6: (2016). DOI: 10.1038/srep36199. Application: Titer Determination.
- Robitaille, K et al. "High-throughput Functional Genomics Identifies Regulators of Primary Human Beta Cell Proliferation" J. Biol. Chem. 291(9):4614-4625 (2016). DOI: 10.1074/jbc.M115.683912 .
- Zhang, M et al. "Epigenetically mediated spontaneous reduction of NFAT1 expression causes imbalanced metabolic activities of articular chondrocytes in aged mice" Osteoarthritis Cartilage. 24(7):1274-1283 (2016). DOI: 10.1016/j.joca.2016.02.003.
- Kurata, M et al. "Using genome-wide CRISPR library screening with library resistant DCK to find new sources of Ara-C drug resistance in AML" Sci. Rep. 6:36199 (2016). DOI: doi:10.1038/srep36199.
- Margul, DJ et al. "Reducing neuroinflammation by delivery of IL-10 encoding lentivirus from multiple-channel bridges" Bioeng Transl Med. 1(2):136-148 (2016). DOI: DOI: 10.1002/btm2.10018.
- Theis, T et al. "Lentiviral Delivery of miR-133b Improves Functional Recovery After Spinal Cord Injury in Mice" Mol Neurobiol. : (2016). DOI: 10.1007/s12035-016-0007-z.
- Henry, A et al. "New role of osteopontin in DNA repair and impact on human glioblastoma radiosensitivity" Oncotarget 7(39):63708-63721 (2016). DOI: 10.18632/oncotarget.11483 .
- Jin, C et al. "Safe engineering of CAR T cells for adoptive cell therapy of cancer using long-term episomal gene transfer" EMBO Mol Med. 8(7):702-711 (2016). DOI: 10.15252/emmm.201505869.
- Bushnell, G. G., Rao, S. S., Hartfield, R. M., Zhang, Y., Oakes, R. S., Jeruss, J. S., & Shea, L. D. "Microporous scaffolds loaded with immunomodulatory lentivirus to study the contribution of immune cell populations to tumor cell recruitment in vivo" Biotechnology and bioengineering. : (2019).
- Johnson, J. O., Chia, R., Brown Jr, R. H., & Landers, J. E. "Mutations in the SPTLC1 gene are a cause of amyotrophic lateral sclerosis that may be amenable to serine supplementation" : (2019).
- Keshwara, R., Hagen, K. R., Abreu-Mota, T., Papaneri, A. B., Liu, D., Wirblich, C., … Schnell, M. J. "A Recombinant Rabies Virus Expressing the Marburg Virus Glycoprotein Is Dependent upon Antibody-Mediated Cellular Cytotoxicity for Protection against Marburg Virus Disease in a Murine Model" Journal of Virology 93(6): (2018). DOI: 10.1128/jvi.01865-18.
- Long, K. L. P.. "The role of oligodendrocytes and myelin in differential susceptibility to stress-induced anxiety (Doctoral Thesis)" University of California Berkeley:United States (2018).
- Rahal, O. M., Nie, L., Chan, L. C., Li, C. W., Hsu, Y. H., Hsu, J., ... & Hung, M. C. "Selective expression of constitutively active pro-apoptotic protein BikDD gene in primary mammary tumors inhibits tumor growth and reduces tumor initiating cells" American journal of cancer research 5(12):3624 (2015).
- Schatoff, E. M., Zafra, M. P., & Dow, L. E. "Base editing the mammalian genome" Methods 164–165:100–108 (2019). DOI: 10.1016/j.ymeth.2019.02.022.
- Scheben, A., Wolter, F., Batley, J., Puchta, H., & Edwards, D. "Towards CRISPR/Cas crops - bringing together genomics and genome editing" New Phytologist 216(3):682–698 (2017). DOI: 10.1111/nph.14702.
- Tsai, . "U" S. Patent Application No.0015473 A1 : (2019).