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Bingo™ CRISPR Point Mutation Cell Line Generation Kit(Basic Version)

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Product Information

Catalog Number	Product Name	Components
EDY003-Y01	Bingo™ CRISPR Point Mutation Cell Line Generation Kit (Basic Version)	Bingo™ Point Mutation Plasmid
EDY003-Y02	Bingo™ CRISPR Point Mutation Cell Line Generation Kit (Advanced Version)	Bingo™ Point Mutation Plasmid
		Cell Lysis Reagents
		PCR Validation Reagents

Description

This kit is the Bingo™ CRISPR Point Mutation Cell Line Generation Kit, designed for the precise and efficient preparation of cell lines with targeted gene point mutations (substitutions, insertions, deletions). It provides cells, plasmids, and key reagents necessary for the gene point mutation process, offering researchers a highly efficient and convenient tool for one-stop construction of point mutation cell lines.

To significantly reduce experimental costs, this kit also includes Cell Lysis and PCR Validation products. Genomic sample preparation takes only 20 minutes and allows downstream PCR experiments without purification. The PCR Validation reagents are compatible with residual cell lysis components in crude genomic samples, enabling rapid and accurate identification of targeted mutation cell genotypes.

Kit Composition

l Bingo™ Point Mutation Plasmid

Component			Specification	Storage Temperature
Helper Plasmids(500 ng/μL)	Bingo^TM PE		50μg	﹣20℃
Target Plasmids(200 ng/μL)	Bingo^TM pegRNA1	Bingo^TMgRNA1	4 μg/pegRNA, 1.5 μg/gRNA	﹣20℃
	Bingo^TM pegRNA2	Bingo^TM gRNA2
	Bingo^TM pegRNA3	Bingo^TM gRNA3
Positive Control(200ng/μL)	Bingo^TM pegRNA Control	Bingo^TM gRNA Control	4 μg/pegRNA, 1.5 μg/gRNA	﹣20℃
Cell (optional)			1×10⁶/tube	Liquid nitrogen

l Cell Lysis Reagents

Component	Specification	Storage Temperature
Buffer A	10 mL	RT
Buffer B	200 μL	-20 ℃

l PCR Validation Reagents

Component	Specification	Storage Temperature
2× High Fidelity Pfu Mix (+Dye)	1.25 mL	-20 ℃
Genotyping Primer F	1 OD Powder	-20 ℃
Genotyping Primer R	1 OD Powder	-20 ℃

Workflow Diagram

Figure 1. Workflow Diagram for Gene Point Mutation

Storage Comditions and Shelf Life

-20°C, valid for 1 year.

FAQ

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

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.

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.

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.

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).