Which of following service are you looking for?
Single gene knockout in a cell line Multiplex gene knockout in a cell line. Specify gene number:_____________
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Name of Target Gene:
NCBI Accession Number (Gene ID):
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Could you please describe what your application is with the KO cell line?
Gene function analysis Assay development Drug screening Bioproduction In vivo tumorigenesis
Other. Please indicate your specific application and requirements:_________________________________
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Comments:
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Do you need GenScript to design gRNAs:
No. Please provide gRNA sequences to be used:______________________________________________________________
Yes. Please specify the gene products:
NCBI accession ID NM or NP: _______________________________________________________________________________
UniProt entry ID: __________________________________________________________________________________________
Sequence to be disrupted:___________________________________________________________________________________
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Does KO of the target gene inhibit cell proliferation?
Yes No Not sure
If yes or no, please provide an evidence to support your choice:____________________________________________________
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Does KO of the target gene inhibit cell survival?
Yes No Not sure
If yes or no, please provide an evidence to support your choice:____________________________________________________
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Name of Target Host Cell Line:___________________________________________
Alternative cell lines:_______________________________________________
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Is the host cell line commercially available?
Yes. Please provide vendor info and Catalog No._________________________________________________________
No.
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Who provides host cell line?
Client (Note: Cell line must be Mycoplasma-free)
GenScript (Note: Only for ATCC cell lines and extra fee and time may be required)
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What is the copy number variation (CNV) of the target gene in the host cell line?
One copy Two copies More. Specify copy number: _________ Not sure
Please provide an evidence to support your choice:____________________________________________________
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Which of following methods do you prefer to enrich the transfected cells?
Antibiotic selection (Blasticidin and/or puromycin) FACS sorting
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What is the suggested method for cell transfection?
Chemical transfection, please specify the reagent:______________________________________
Electroporation, please specify the program:___________________________________________
Nucleofection, please specify the kit and program:______________________________________
Lentivirus
Provide transfection efficiency for the suggested method if possible:____________________________________
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Which of following promoters work best in the host cell line?
CMV CBh EF1-a Not sure
if it is not listed above, please specify: ________________________
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Growth condition of host cell line? Adherent Suspension Both
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What are the medium and additives for cell growth?
Medium:_____________________________________________ Additives:__________________________________________________
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Cell subculture protocol:
The cell line is passaged: ___________times per week in a ratio: ______________.
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Do you need GenScript to follow any special cell culture routine? Yes, see below No
Please provide the protocol with information about the cell line and any special growth characteristics or requirements:
__________________________________________________________________________________________
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Is the cell line immortalized? Yes No Not sure
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Does serial dilution affect cell growth rate? Yes No Not sure
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Do the cells contain any human pathogen that may be harmful to human?
Yes, please specify:___________________________ No
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Which type of deliverables are you looking for:
I just need CRISPR knockout cell pools
I need CRISPR knockout cell lines
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Which of following genotype of knockout cell line do you want (Validated by genome DNA sequencing):
Single allele knockout Bi-allelic knockout All-alleles knockout (Allele number is unknown)
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How many knockout clones do you want (Extra fees may be charged):
One clone Two clones Three clones More clones, specify the amount:_________________
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Please indicate the preferred type of optional analysis to validate transgenic cell line (Extra fees may be charged):
Reverse-transcribe (RT) PCR (Verify the INDELs by sequencing at mRNA levels)
Western blot (Validated antibodies to be provided by customer)
FACS analysis (Validated antibodies to be provided by customer)
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Please indicate the preferred type of optional analysis to characterize transgenic cell line (Extra fees may be charged):
Off-target analysis Others, please specify:_________________
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