BacPAK qPCR Titration Kit
BacPAK qPCR Titration Kit
Brand: Takara Bio.
In stock
SKU
BacPAK qPCR Titration Kit
This kit can be used along with real-time PCR equipment to rapidly quantify the virion content of baculoviral supernatants. The copy numbers of viral DNA genomes contained in virus samples are determined by comparing the qPCR threshold cycles (Ct) derived from serial dilutions of purified viral DNA to the Ct values derived from dilutions of a calibrated control template. The kit includes a viral nucleic acid purification kit, a calibrated BacPAK DNA Control Template, primers, and all the reagents required to conduct 200 qPCR reactions using either the BacPAK DNA Control Template or purified viral DNA samples.
Overview
- Saves time by shortening baculovirus expression experiments up to six days
- Eliminates troublesome plaque assays
- Compatible with any AcMNPV-based baculovirus expression systems
- Rapid determination of viral titers
- Harvest, titer, and infect in a single day
- Accurate titers produce the most consistent infection results
Applications
- Determine baculovirus titers, in order to optimize recombinant protein expression
The BacPAK qPCR Titration kit offers a wide dynamic range
The BacPAK qPCR Titration kit offers a wide dynamic range. The BacPAK DNA Control Template was serially diluted from 108–103 copies per sample and analyzed with the BacPAK qPCR Titration Kit. The amplification plots (Panel A) show a dynamic range of at least six orders of magnitude with no NTC (No-Template Control) background. The standard curve (Panel B) obtained by plotting the Ct values (determined from the amplification plots in Panel A) against the log of the DNA copy number in each sample, demonstrates a strong linear correlation between the Ct and the DNA copy number (log scale), with R2 = 1.000 and a PCR efficiency of 100.0.07.
The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System
The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System. A PCR fragment containing your gene of interest is simultaneously and directly cloned into the In-Fusion Ready BacPAK Vector pair to generate N- and C-terminal-tagged constructs. Verified clones are then cotransfected with BacPAK6 viral DNA into insect cells to initiate recombination via the AcMNPV sequences, and replication of recombinant virus. Following virus amplification, infection of insect cell cultures produces recombinant protein, which is then purified by TALON technology.
Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1). An In-Fusion Ready BacPAK vector was used to generate a recombinant baculovirus harboring an N-terminal tagged expression construct for the Living Colors fluorescent protein, AcGFP1. Sf9 cells infected with the virus expressed high levels of AcGFP1 and became highly fluorescent. Analysis by flow cytometry revealed that the mean fluorescence intensity of the infected cells was approximately 440-fold greater than that of the uninfected control.
Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1). In-Fusion Ready BacPAK vectors were used to generate recombinant baculoviruses harboring N- or C-terminal-tagged AcGFP1 expression constructs. Insect cells (Sf9) infected with either virus express AcGFP1 and thus appear green when viewed by fluorescence microscopy. Panel A. BacPAK-Nterm-6xHN-AcGFP1. Panel B. BacPAK-Cterm- 6xHN-AcGFP1
The BacPAK qPCR Titration Protocol
The BacPAK qPCR Titration Protocol.
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The BacPAK qPCR Titration kit offers a wide dynamic range
The BacPAK qPCR Titration kit offers a wide dynamic range. The BacPAK DNA Control Template was serially diluted from 108–103 copies per sample and analyzed with the BacPAK qPCR Titration Kit. The amplification plots (Panel A) show a dynamic range of at least six orders of magnitude with no NTC (No-Template Control) background. The standard curve (Panel B) obtained by plotting the Ct values (determined from the amplification plots in Panel A) against the log of the DNA copy number in each sample, demonstrates a strong linear correlation between the Ct and the DNA copy number (log scale), with R2 = 1.000 and a PCR efficiency of 100.0.07.
The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System
The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System. A PCR fragment containing your gene of interest is simultaneously and directly cloned into the In-Fusion Ready BacPAK Vector pair to generate N- and C-terminal-tagged constructs. Verified clones are then cotransfected with BacPAK6 viral DNA into insect cells to initiate recombination via the AcMNPV sequences, and replication of recombinant virus. Following virus amplification, infection of insect cell cultures produces recombinant protein, which is then purified by TALON technology.
Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1). An In-Fusion Ready BacPAK vector was used to generate a recombinant baculovirus harboring an N-terminal tagged expression construct for the Living Colors fluorescent protein, AcGFP1. Sf9 cells infected with the virus expressed high levels of AcGFP1 and became highly fluorescent. Analysis by flow cytometry revealed that the mean fluorescence intensity of the infected cells was approximately 440-fold greater than that of the uninfected control.
Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1). In-Fusion Ready BacPAK vectors were used to generate recombinant baculoviruses harboring N- or C-terminal-tagged AcGFP1 expression constructs. Insect cells (Sf9) infected with either virus express AcGFP1 and thus appear green when viewed by fluorescence microscopy. Panel A. BacPAK-Nterm-6xHN-AcGFP1. Panel B. BacPAK-Cterm- 6xHN-AcGFP1
The BacPAK qPCR Titration Protocol
The BacPAK qPCR Titration Protocol.
