PiggyBac qPCR Copy Number Kit
Products
| Catalog Number | Description | Size | Price | Quantity | Add to Cart | |||
|---|---|---|---|---|---|---|---|---|
| PBC100A-1 | piggyBac qPCR copy number kit, 20 reactions | 20 Reactions | $285 |
|
||||
Overview
Overview
Confidently count your PiggyBac insertsWhen 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:
- Calculate the average Cts for the PiggyBac inserts and for the UCR1 loci (there are two UCR1 elements per genome)
- 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:
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.
FAQs
Resources
Related Products
Citations
-
Brouwer, I, de Kort, MAC & Lenstra, TL. (2024) Measuring Transcription Dynamics of Individual Genes Inside Living Cells. Methods in molecular biology (Clifton, N.J.). 2024; 2694:235-265. PM ID: 37824008
-
Matta, SK, et al. (2024) Genome-wide and targeted CRISPR screens identify RNF213 as a mediator of interferon gamma-dependent pathogen restriction in human cells. Proceedings of the National Academy of Sciences of the United States of America. 2024; 121(1):e2315865120. PM ID: 38147552
-
Cho, MG, et al. (2024) MRE11 liberates cGAS from nucleosome sequestration during tumorigenesis. Nature. 2024; 625(7995):585-592. PM ID: 38200309
-
Du, M, et al. (2024) Direct observation of a condensate effect on super-enhancer controlled gene bursting. Cell. 2024; 187(2):331-344.e17. PM ID: 38194964
-
Schmitt, J, et al. (2024) Repurposing an endogenous degradation domain for antibody-mediated disposal of cell-surface proteins. EMBO reports. 2024;. PM ID: 38287192
-
Byrnes, AE, et al. (2024) A fluorescent splice-switching mouse model enables high-throughput, sensitive quantification of antisense oligonucleotide delivery and activity. Cell reports methods. 2024; 4(1):100673. PM ID: 38171361
-
Daiki, K, et al. (2024) Blood Endocan as a Biomarker for Breast Cancer Recurrence. Preprint. 2024;. Link: Preprint
-
Koeppel, J, et al. (2024) Randomizing the human genome by engineering recombination between repeat elements. bioRxiv. 2024;. Link: bioRxiv
-
Kortleve, D, et al. (2024) TCR-engineered T-cells directed against Ropporin-1 constitute a safe and effective treatment for triple-negative breast cancer in near-clinical models. bioRxiv. 2024;. Link: bioRxiv
-
Haakonsen, DL, et al. (2024) Stress response silencing by an E3 ligase mutated in neurodegeneration. Nature. 2024; 626(8000):874-880. PM ID: 38297121
-
Gupta, P, et al. (2024) Development of pathophysiologically relevant models of sickle cell disease and β-thalassemia for therapeutic studies. Nature communications. 2024; 15(1):1794. PM ID: 38413594
-
Company, C, et al. (2024) Logical design of synthetic cis-regulatory DNA for genetic tracing of cell identities and state changes. Nature communications. 2024; 15(1):897. PM ID: 38316783
-
Yang, L, et al. (2024) Uncovering receptor-ligand interactions using a high-avidity CRISPR activation screening platform. Science advances. 2024; 10(7):eadj2445. PM ID: 38354234
-
Kubara, K, et al. (2024) Lymph node macrophages drive innate immune responses to enhance the anti-tumor efficacy of mRNA vaccines. Molecular therapy : the journal of the American Society of Gene Therapy. 2024;. PM ID: 38243602
-
Ng-Blichfeldt, J, et al. (2024) Identification of a core transcriptional program driving the human renal mesenchymal-to-epithelial transition. Developmental Cell. 2024;. Link: Developmental Cell
-
Yang, J, Cook, L & Chen, Z. (2024) Systematic evaluation of retroviral LTRs as cis-regulatory elements in mouse embryos. Cell reports. 2024; 43(3):113775. PM ID: 38381606
-
Taglini, F, et al. (2024) DNMT3B PWWP mutations cause hypermethylation of heterochromatin. EMBO reports. 2024;. PM ID: 38291337
-
Tanase-Nakao, K, et al. (2024) Genotype-Phenotype Correlations in Thirty Japanese Patients with Congenital Hypothyroidism Attributable to TG Defects. The Journal of clinical endocrinology and metabolism. 2024;. PM ID: 38373250
-
Alsouri, S, et al. (2024) Actinin-4 controls survival signaling in B cells by limiting the lateral mobility of B-cell antigen receptors. European journal of immunology. 2024;:e2350774. PM ID: 38299456
-
Ke, X, et al. (2024) Establishment of a novel minigenome system for the identification of drugs targeting Nipah virus replication. The Journal of general virology. 2024; 105(1). PM ID: 38180473
- See More
Products
| Catalog Number | Description | Size | Price | Quantity | Add to Cart | |||
|---|---|---|---|---|---|---|---|---|
| PBC100A-1 | piggyBac qPCR copy number kit, 20 reactions | 20 Reactions | $285 |
|
||||
Overview
Overview
Confidently count your PiggyBac insertsWhen 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:
- Calculate the average Cts for the PiggyBac inserts and for the UCR1 loci (there are two UCR1 elements per genome)
- 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:
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.
FAQs
Citations
-
Brouwer, I, de Kort, MAC & Lenstra, TL. (2024) Measuring Transcription Dynamics of Individual Genes Inside Living Cells. Methods in molecular biology (Clifton, N.J.). 2024; 2694:235-265. PM ID: 37824008
-
Matta, SK, et al. (2024) Genome-wide and targeted CRISPR screens identify RNF213 as a mediator of interferon gamma-dependent pathogen restriction in human cells. Proceedings of the National Academy of Sciences of the United States of America. 2024; 121(1):e2315865120. PM ID: 38147552
-
Cho, MG, et al. (2024) MRE11 liberates cGAS from nucleosome sequestration during tumorigenesis. Nature. 2024; 625(7995):585-592. PM ID: 38200309
-
Du, M, et al. (2024) Direct observation of a condensate effect on super-enhancer controlled gene bursting. Cell. 2024; 187(2):331-344.e17. PM ID: 38194964
-
Schmitt, J, et al. (2024) Repurposing an endogenous degradation domain for antibody-mediated disposal of cell-surface proteins. EMBO reports. 2024;. PM ID: 38287192
-
Byrnes, AE, et al. (2024) A fluorescent splice-switching mouse model enables high-throughput, sensitive quantification of antisense oligonucleotide delivery and activity. Cell reports methods. 2024; 4(1):100673. PM ID: 38171361
-
Daiki, K, et al. (2024) Blood Endocan as a Biomarker for Breast Cancer Recurrence. Preprint. 2024;. Link: Preprint
-
Koeppel, J, et al. (2024) Randomizing the human genome by engineering recombination between repeat elements. bioRxiv. 2024;. Link: bioRxiv
-
Kortleve, D, et al. (2024) TCR-engineered T-cells directed against Ropporin-1 constitute a safe and effective treatment for triple-negative breast cancer in near-clinical models. bioRxiv. 2024;. Link: bioRxiv
-
Haakonsen, DL, et al. (2024) Stress response silencing by an E3 ligase mutated in neurodegeneration. Nature. 2024; 626(8000):874-880. PM ID: 38297121
-
Gupta, P, et al. (2024) Development of pathophysiologically relevant models of sickle cell disease and β-thalassemia for therapeutic studies. Nature communications. 2024; 15(1):1794. PM ID: 38413594
-
Company, C, et al. (2024) Logical design of synthetic cis-regulatory DNA for genetic tracing of cell identities and state changes. Nature communications. 2024; 15(1):897. PM ID: 38316783
-
Yang, L, et al. (2024) Uncovering receptor-ligand interactions using a high-avidity CRISPR activation screening platform. Science advances. 2024; 10(7):eadj2445. PM ID: 38354234
-
Kubara, K, et al. (2024) Lymph node macrophages drive innate immune responses to enhance the anti-tumor efficacy of mRNA vaccines. Molecular therapy : the journal of the American Society of Gene Therapy. 2024;. PM ID: 38243602
-
Ng-Blichfeldt, J, et al. (2024) Identification of a core transcriptional program driving the human renal mesenchymal-to-epithelial transition. Developmental Cell. 2024;. Link: Developmental Cell
-
Yang, J, Cook, L & Chen, Z. (2024) Systematic evaluation of retroviral LTRs as cis-regulatory elements in mouse embryos. Cell reports. 2024; 43(3):113775. PM ID: 38381606
-
Taglini, F, et al. (2024) DNMT3B PWWP mutations cause hypermethylation of heterochromatin. EMBO reports. 2024;. PM ID: 38291337
-
Tanase-Nakao, K, et al. (2024) Genotype-Phenotype Correlations in Thirty Japanese Patients with Congenital Hypothyroidism Attributable to TG Defects. The Journal of clinical endocrinology and metabolism. 2024;. PM ID: 38373250
-
Alsouri, S, et al. (2024) Actinin-4 controls survival signaling in B cells by limiting the lateral mobility of B-cell antigen receptors. European journal of immunology. 2024;:e2350774. PM ID: 38299456
-
Ke, X, et al. (2024) Establishment of a novel minigenome system for the identification of drugs targeting Nipah virus replication. The Journal of general virology. 2024; 105(1). PM ID: 38180473
- See More