Plasmid vector

... try to ligate annealed oligos into double digested (different enzymes) plasmid. The oligos I annealed were not modified with phosphates. Do I need ... it is not necessary to dephosphorylate my double-digested open vector as long as it was cut with two different enzymes. Thus, the 5'-end ...

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... issue (and difference between the two) are not so obvious for us. Our vector is a circular plasmid. Can you elaborate on what you mean by two linear pieces that are homologous to each ...

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... way to make negative control is to set up a ligation of your digested vector with water (or TE buffer/elution buffer you are using) in place of ... not work anymore. The other suggestion - transformation using wrong plasmid (another antibiotic resistance) - is also wrong and has no value.

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The vector is a circular molecule containing a total of 5730 bp. It has sites for the restriction ... cut with PstI and EcoRI. (The chicken gene fragment replacing the smaller segment of plasmid DNA between these restriction enzyme sites). Where is says Pstl, isn't it meant to ...

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1-are you sure you have an insert in your vector? If not, there is a big chance the vector is empty and therefore you have ... fluorescent dye I dont know what can you see on your gel. When you load uncut plasmid and cut plasmid on the gel into separate wells and run electrophoresis-how ...

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