Chloroquine transfection protocol

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  1. aksakal XenForo Moderator

    Chloroquine transfection protocol


    This procedure can be modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used for a variety of downstream applications such as stable-cell line generation.

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    If chloroquine is not used, it is unnecessary to change the medium prior to transfection. On some occasions, we have obtained slightly improved transfection efficiencies by adding the chloroquine to a 11 mixture of 293 conditioned media obtained from any of the 293-based cell lines and fresh GM. Troubleshooting Transfection Experiments. Transfection Protocol Calculator › Low transfection efficiency. Do not use antibiotics such as chloroquine, penicillin, or streptomycin in growth medium because during transfection, cells are more permeable to antibiotics, which may cause toxicity. Phoenix - Helper free protocol. The following protocol is an adaptation of a review in press, it has been changed to accomodate the Phoenix systems protocol developed by Achacoso and Nolan. This is one of four transfection protocols you can find in this web page set.

    Test a variety of brands and lots of FBS to find one suitable with your protocols. Last Upload: August 26, 2016 *Pro-Tip* Different brands and lots of FBS can promote or inhibit transfection.

    Chloroquine transfection protocol

    Helper Dependent Protocol - Stanford University, Troubleshooting Transfection Experiments Thermo Fisher.

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  6. This protocol is a modified version of a published method 4, in which calcium phosphate–based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells.

    • Calcium phosphate–mediated transfection of eukaryotic..
    • Phoenix - Helper free protocol - Stanford University.
    • Addgene Lentivirus Production Protocol.

    Transfection Experiment Transfection Efficacy Reagent Volume Tissue Culture Grade Chloroquine Diphosphate These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves. Since the publication of the original method1, increases in efficiency have been achieved by including additional steps such as glycerol shock2 and/or chloroquine treatment3. This protocol is a modified version of a published method4, in which calcium phosphate–based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells were rigorously optimized. Within 5-10’ of adding the chloroquine. 7. For 15cm plates, add 1.69ml of the transfection buffer to the DNA-CaCl2 solution by “bubbling”. To “bubble”, use two mechanical pipeters Pipet-AidTM, one with a 1ml pipette to bubble air into the DNA-CaCl2 solution and the other to slowly drip the transfection buffer into the conical tube.

     
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