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FAQ
Inconsistent results with repeated testing
PCR detection of Mycoplasma is highly sensitive, so avoid cross-contamination between samples. Ensure sterile conditions when handling each sample individually. After sample processing and PCR reaction, allow samples to cool before proceeding to the next step to avoid aerosol contamination.
Can the kit detect Mycoplasma in reagents?
The kit can detect Mycoplasma in common cell culture reagents, such as media and serum, without the need for sample preparation. Simply proceed directly to PCR reaction and gel loading. However, the kit cannot detect Mycoplasma in organic solvents like DMSO or ethanol.
Bands appear in the negative control
Use a freshly opened negative control for retesting. Ensure to change pipette tips during sample loading and prioritize loading the negative control first to avoid cross-contamination between samples.
Inconsistent results with repeated testing
PCR detection of Mycoplasma is highly sensitive, so avoid cross-contamination between samples. Ensure sterile conditions when handling each sample individually. After sample processing and PCR reaction, allow samples to cool before proceeding to the next step to avoid aerosol contamination.
Can the kit detect Mycoplasma in reagents?
The kit can detect Mycoplasma in common cell culture reagents, such as media and serum, without the need for sample preparation. Simply proceed directly to PCR reaction and gel loading. However, the kit cannot detect Mycoplasma in organic solvents like DMSO or ethanol.
Does the Bingo™ CRISPR Point Mutation Cell Line Generation Kit recommend lipofection or electroporation?
The Bingo™ PE, pegRNA, and gRNA in the kit are all plasmids, compatible with both lipofection and electroporation. You can choose the appropriate transfection method and parameters based on cell type. Generally, adherent cells can be transfected by either method, while therapeutically relevant cells are recommended to use electroporation
Can cells constructed with the Bingo™ CRISPR Point Mutation Cell Line Generation Kit be stably passaged?
Yes. The Bingo™ CRISPR Point Mutation Cell Line Generation Kit targets genomic DNA for gene mutation, allowing the genotype of the target cells to be stably inherited by subsequent generations.
How can I verify the editing activity of the Bingo™ CRISPR Point Mutation Cell Line Generation Kit?
The kit’s Positive Control has been validated in HeLa cells (human gene) and N2a cells (mouse gene). Due to the high heterogeneity of cells, transfection and editing efficiency may vary in different cell types using the same kit.
Why do cells plated in a 96-well plate for single-clone screening grow slowly or die, even though single-clone formation was good in the pre-experiment?
Gene mutations may affect cell viability. Before conducting a gene point mutation experiment, it is advisable to consult relevant literature to understand the function of the target gene. If the target gene plays a critical role in cell proliferation or survival, point mutations may hinder these processes, making it difficult to obtain positive cells.
What should I do if the transfection efficiency in target cells is low?
It is recommended to conduct a pre-transfection experiment before the main experiment. Try different transfection methods to find optimal conditions, such as common chemical transfection methods (e.g., lipofection) or physical methods (e.g., electroporation).
Does the piggyBac Transposon System Kit recommend lipofection or electroporation?
The PB system in this kit includes plasmids compatible with lipofection, electroporation, or other transfection methods. You can select the transfection method and parameters based on cell type. Most adherent cells are compatible with various transfection methods, while electroporation is recommended for therapeutically relevant cells.
What should I do if the transfection efficiency in target cells is low?
Before the main experiment, conduct a transfection pre-experiment to explore different transfection methods and optimize conditions. Efficient transfection is essential for successful gene transposition.
How can I verify the transposition activity of the piggyBac Transposon System Kit?
The transposon plasmid in this kit contains the CopGFP gene. Successfully transposed cells will exhibit green fluorescence under a fluorescence microscope. Due to cell heterogeneity, transfection and transposition efficiencies may vary across cell types. To minimize transposition variability, optimize transfection methods for target cells.
Can cells constructed with the piggyBac Transposon System Kit be stably passaged?
Yes. The piggyBac Transposon System Kit targets genomic DNA for gene transposition, allowing the genotype of target cells to be stably inherited.
Note: Maintain transposed cells with half-dose puromycin for stability.
Are all types of genes suitable for KO cell lines?
Not all genes are suitable for knockout. Some gene knockouts may result in cell death or severe dysfunction, particularly for essential genes. In such cases, conditional knockouts or gene knockdowns (e.g., RNAi) may be used instead.
