PiggyBac qPCR Copy Number Kit

Find out how many copies of your PiggyBac insert have been integrated into the genome with this accurate, qPCR-based kit

Products

Catalog Number Description Size Price Quantity Add to Cart
PBC100A-1 piggyBac qPCR copy number kit, 20 reactions   20 Reactions $270
- +

Overview

Overview

Confidently count your PiggyBac inserts

When you need to know how many PiggyBac integration events have happened, turn to SBI’s PiggyBac qPCR Copy Number Kit. This kit uses qPCR to measure the number of PiggyBac inserts relative to a specific genomic locus—the UCR1 element—with the PiggyBac insert copy number calculated using the cycle threshold (Ct) values of the UCR1 signal relative to the PiggyBac insert signal.

The PiggyBac qPCR Copy Number Kit comes with enough reagents—UCR1 primers, PiggyBac primers, and cell lysis buffer—for twenty copy number determinations, and is compatible with all of SBI’s PiggyBac Vectors.

NOTE: Your cells must be passaged at least once before performing this copy number measurement to ensure that residual, non-integrated piggyBac transposon plasmid does not interfere with the qPCR reaction.

How It Works

How It Works

Calculate PiggyBac insert copy number

The PiggyBac qPCR Copy Number Kit provides robust PiggyBac insert copy number determination. To calculate the PiggyBac insert copy number from the Ct values:

  1. Calculate the average Cts for the PiggyBac inserts and for the UCR1 loci (there are two UCR1 elements per genome)
  2. The copy number is the ΔΔCt/2—the ΔΔCt value must be divided by two to account for the two UCR1 elements:

ΔΔCt = 2-((average PiggyBac insert Ct) – (average UCR1 Ct))
PiggyBac insert copy number = ΔΔCt/2

You can change the number of PiggyBac insertions by adjusting the ratio of PiggyBac Vector to Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1).

Supporting Data

Supporting Data

Robust PiggyBac insert copy number determination

Example PiggyBac qPCR Copy Number Kit data from a PiggyBac Vector titration study—the PiggyBac insert copy number can be changed by adjusting the ratio of PiggyBac Vector to Super PiggyBac Transposase Expression Vector:

Adjust the PiggyBac insert copy number by changing the ratio of PiggyBac Vector to Super PiggyBac Transposase

Figure 1. Adjust the PiggyBac insert copy number by changing the ratio of PiggyBac Vector to Super PiggyBac Transposase. To achieve the indicated ratio of PiggyBac Vector to Super PiggyBac Transposase Expression Vector, amount of PiggyBac Vector added was 100 ng, 300 ng, 500 ng, 700 ng, and 1,000 ng, respectively, while the amount of Super PiggyBac Transposase Expression Vector was held constant at 100 ng.

Resources

Citations

  • Uchino, S, et al. (2022) Live imaging of transcription sites using an elongating RNA polymerase II-specific probe. The Journal of cell biology. 1970 Jan 1; 221(2). PM ID: 34854870
  • Teixeira, A, et al. (2022) CelloSelect – A synthetic cellobiose metabolic pathway for selection of stable transgenic CHO cell lines. Metabolic Engineering. 1970 Jan 1; 70:23-30. Link: Metabolic Engineering
  • Rui, Y, et al. (2022) High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA. Science advances. 1970 Jan 1; 8(1):eabk2855. PM ID: 34985952
  • Vásquez-Limeta, A, et al. (2022) CPAP insufficiency leads to incomplete centrioles that duplicate but fragment. The Journal of cell biology. 1970 Jan 1; 221(5). PM ID: 35404385
  • Su, CJ, et al. (2022) Ligand-receptor promiscuity enables cellular addressing. Cell systems. 1970 Jan 1;. PM ID: 35421362
  • Klumpe, HE, et al. (2022) The context-dependent, combinatorial logic of BMP signaling. Cell systems. 1970 Jan 1;. PM ID: 35421361
  • Kitano, H, Kawabe, Y & Kamihira, M. (2022) HepG2-Based Designer Cells with Heat-Inducible Enhanced Liver Functions. Cells. 1970 Jan 1; 11(7). PM ID: 35406758
  • Sugiman-Marangos, SN, et al. (2022) Structures of distant diphtheria toxin homologs reveal functional determinants of an evolutionarily conserved toxin scaffold. Communications biology. 1970 Jan 1; 5(1):375. PM ID: 35440624
  • Ma, X, et al. (2022) Validation of reliable safe harbor locus for efficient porcine transgenesis. Functional & integrative genomics. 1970 Jan 1;. PM ID: 35412198
  • Nishimura, K, et al. (2022) Rapid conversion of human induced pluripotent stem cells into dopaminergic neurons by inducible expression of two transcription factors. Stem cells and development. 1970 Jan 1;. PM ID: 35420042
  • Liu, Z, Ramirez, A & Liu, X. (2022) Live Cell Imaging of Spatiotemporal Ca2+ Fluctuation Responses to Anticancer Drugs. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2488:227-236. PM ID: 35347692
  • Dandridge, S. (2022) Honors Thesis: Defining the effect of zinc on the proliferation of MDA-MB-231 cells compared to MCF10A cells. Thesis. 1970 Jan 1;. Link: Thesis
  • Yang, D, et al. (2022) Lineage tracing reveals the phylodynamics, plasticity, and paths of tumor evolution. Cell. 1970 Jan 1; 185(11):1905-1923.e25. PM ID: 35523183
  • Biswas, S, et al. (2022) Long-term hepatitis B virus infection of rhesus macaques requires suppression of host immunity. Nature communications. 1970 Jan 1; 13(1):2995. PM ID: 35637225
  • Breau, KA, et al. (2022) Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells. Stem cell reports. 1970 Jan 1;. PM ID: 35523179
  • Lensch, S, et al. (2022) Dynamic spreading of chromatin-mediated gene silencing and reactivation between neighboring genes in single cells. eLife. 1970 Jan 1; 11. PM ID: 35678392
  • Gu, J, Sumer, H & Cromer, B. (2022) Efficient Generation of Stable Cell Lines with Inducible Neuronal Transgene Expression Using the piggyBac Transposon System. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2495:49-66. PM ID: 35696027
  • Wang, S, et al. (2021) Budding epithelial morphogenesis driven by cell-matrix versus cell-cell adhesion. Cell. 1970 Jan 1;. PM ID: 34133940
  • Ng, YH, et al. (2021) Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics. Stem cell reports. 1970 Jan 1;. PM ID: 34171286
  • Ukaji, T, et al. (2021) Novel knock-in mouse model for the evaluation of the therapeutic efficacy and toxicity of human podoplanin-targeting agents. Cancer science. 1970 Jan 1; 112(6):2299-2313. PM ID: 33735501

Products

Catalog Number Description Size Price Quantity Add to Cart
PBC100A-1 piggyBac qPCR copy number kit, 20 reactions   20 Reactions $270
- +

Overview

Overview

Confidently count your PiggyBac inserts

When you need to know how many PiggyBac integration events have happened, turn to SBI’s PiggyBac qPCR Copy Number Kit. This kit uses qPCR to measure the number of PiggyBac inserts relative to a specific genomic locus—the UCR1 element—with the PiggyBac insert copy number calculated using the cycle threshold (Ct) values of the UCR1 signal relative to the PiggyBac insert signal.

The PiggyBac qPCR Copy Number Kit comes with enough reagents—UCR1 primers, PiggyBac primers, and cell lysis buffer—for twenty copy number determinations, and is compatible with all of SBI’s PiggyBac Vectors.

NOTE: Your cells must be passaged at least once before performing this copy number measurement to ensure that residual, non-integrated piggyBac transposon plasmid does not interfere with the qPCR reaction.

How It Works

How It Works

Calculate PiggyBac insert copy number

The PiggyBac qPCR Copy Number Kit provides robust PiggyBac insert copy number determination. To calculate the PiggyBac insert copy number from the Ct values:

  1. Calculate the average Cts for the PiggyBac inserts and for the UCR1 loci (there are two UCR1 elements per genome)
  2. The copy number is the ΔΔCt/2—the ΔΔCt value must be divided by two to account for the two UCR1 elements:

ΔΔCt = 2-((average PiggyBac insert Ct) – (average UCR1 Ct))
PiggyBac insert copy number = ΔΔCt/2

You can change the number of PiggyBac insertions by adjusting the ratio of PiggyBac Vector to Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1).

Supporting Data

Supporting Data

Robust PiggyBac insert copy number determination

Example PiggyBac qPCR Copy Number Kit data from a PiggyBac Vector titration study—the PiggyBac insert copy number can be changed by adjusting the ratio of PiggyBac Vector to Super PiggyBac Transposase Expression Vector:

Adjust the PiggyBac insert copy number by changing the ratio of PiggyBac Vector to Super PiggyBac Transposase

Figure 1. Adjust the PiggyBac insert copy number by changing the ratio of PiggyBac Vector to Super PiggyBac Transposase. To achieve the indicated ratio of PiggyBac Vector to Super PiggyBac Transposase Expression Vector, amount of PiggyBac Vector added was 100 ng, 300 ng, 500 ng, 700 ng, and 1,000 ng, respectively, while the amount of Super PiggyBac Transposase Expression Vector was held constant at 100 ng.

Citations

  • Uchino, S, et al. (2022) Live imaging of transcription sites using an elongating RNA polymerase II-specific probe. The Journal of cell biology. 1970 Jan 1; 221(2). PM ID: 34854870
  • Teixeira, A, et al. (2022) CelloSelect – A synthetic cellobiose metabolic pathway for selection of stable transgenic CHO cell lines. Metabolic Engineering. 1970 Jan 1; 70:23-30. Link: Metabolic Engineering
  • Rui, Y, et al. (2022) High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA. Science advances. 1970 Jan 1; 8(1):eabk2855. PM ID: 34985952
  • Vásquez-Limeta, A, et al. (2022) CPAP insufficiency leads to incomplete centrioles that duplicate but fragment. The Journal of cell biology. 1970 Jan 1; 221(5). PM ID: 35404385
  • Su, CJ, et al. (2022) Ligand-receptor promiscuity enables cellular addressing. Cell systems. 1970 Jan 1;. PM ID: 35421362
  • Klumpe, HE, et al. (2022) The context-dependent, combinatorial logic of BMP signaling. Cell systems. 1970 Jan 1;. PM ID: 35421361
  • Kitano, H, Kawabe, Y & Kamihira, M. (2022) HepG2-Based Designer Cells with Heat-Inducible Enhanced Liver Functions. Cells. 1970 Jan 1; 11(7). PM ID: 35406758
  • Sugiman-Marangos, SN, et al. (2022) Structures of distant diphtheria toxin homologs reveal functional determinants of an evolutionarily conserved toxin scaffold. Communications biology. 1970 Jan 1; 5(1):375. PM ID: 35440624
  • Ma, X, et al. (2022) Validation of reliable safe harbor locus for efficient porcine transgenesis. Functional & integrative genomics. 1970 Jan 1;. PM ID: 35412198
  • Nishimura, K, et al. (2022) Rapid conversion of human induced pluripotent stem cells into dopaminergic neurons by inducible expression of two transcription factors. Stem cells and development. 1970 Jan 1;. PM ID: 35420042
  • Liu, Z, Ramirez, A & Liu, X. (2022) Live Cell Imaging of Spatiotemporal Ca2+ Fluctuation Responses to Anticancer Drugs. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2488:227-236. PM ID: 35347692
  • Dandridge, S. (2022) Honors Thesis: Defining the effect of zinc on the proliferation of MDA-MB-231 cells compared to MCF10A cells. Thesis. 1970 Jan 1;. Link: Thesis
  • Yang, D, et al. (2022) Lineage tracing reveals the phylodynamics, plasticity, and paths of tumor evolution. Cell. 1970 Jan 1; 185(11):1905-1923.e25. PM ID: 35523183
  • Biswas, S, et al. (2022) Long-term hepatitis B virus infection of rhesus macaques requires suppression of host immunity. Nature communications. 1970 Jan 1; 13(1):2995. PM ID: 35637225
  • Breau, KA, et al. (2022) Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells. Stem cell reports. 1970 Jan 1;. PM ID: 35523179
  • Lensch, S, et al. (2022) Dynamic spreading of chromatin-mediated gene silencing and reactivation between neighboring genes in single cells. eLife. 1970 Jan 1; 11. PM ID: 35678392
  • Gu, J, Sumer, H & Cromer, B. (2022) Efficient Generation of Stable Cell Lines with Inducible Neuronal Transgene Expression Using the piggyBac Transposon System. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2495:49-66. PM ID: 35696027
  • Wang, S, et al. (2021) Budding epithelial morphogenesis driven by cell-matrix versus cell-cell adhesion. Cell. 1970 Jan 1;. PM ID: 34133940
  • Ng, YH, et al. (2021) Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics. Stem cell reports. 1970 Jan 1;. PM ID: 34171286
  • Ukaji, T, et al. (2021) Novel knock-in mouse model for the evaluation of the therapeutic efficacy and toxicity of human podoplanin-targeting agents. Cancer science. 1970 Jan 1; 112(6):2299-2313. PM ID: 33735501