I want to confirm my transformed construct in pEarleyGate 201 and 103, but first I want to check the plasmid on gel. what should be optimum amount to load
100 ng is absolutely enough. It is better not to be overloaded ... Of course it depends also on the well/gel size.
You should be able to easy visualize 300ng of plasmid, in a gel stained with EtBr or SYBR. I usually load 500-1000ng, just to be safe.
100 ng is absolutely enough. It is better not to be overloaded ... Of course it depends also on the well/gel size.
Use an amount that is comparable with your ladder mass on an agarose gel, eg. use at least 50ng/microliter DNA for 50ng/microliter ladder. Note: make sure linearize your plasmid before running for gel analysis
Firstly, You should use to UV spectrophotometric for measurement of DNA concentration and purity. Then, use at least 100 ng/microliter DNA for running gel analysis
Thanks alot Wael Bahnan, Goritsa Rakleova, Hung king Tiong and Luan Luong Chu for nice suggestions
@ Hung King Tiong, "ng/uL" mentioned is a unit of concentration, it won't give readers much info. One should use 'ng' as the unit for how much DNA. For example, for 100 ng/microliter DNA stock, if 1 uL is used, there will be 100 ng DNA; if 2 uL is used, then there is 200 ng DNA.
It also depends on which staining agent you are using. Some are more sensitive than others. For example: whether the EtBr or the GelRed ("a non-toxic staining agent") [see attached picture: lanes with 200 ng, 100 ng, 50 ng, 25 ng DNA].
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