Often circular segments of DNA that can replicate outside of the chromosomes. Generally found in bacteria.
Often circular segments of DNA that can replicate outside of the chromosomes. Generally found in bacteria.
Questions tagged [plasmids]
134 questions
28
votes
6 answers
How should I ship plasmids?
I shipped 10 µL of my vector miniprep to a collaborator in a 1.5 mL eppendorf parafilmed shut and stuffed into a 50 mL conical with some paper-towel padding. However, something happened on the way and there was nothing (no liquid) in the tube when…
Nick T
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8
votes
1 answer
Taxonomy for plasmids
I liked the question about F' and R plasmids.
What is a good resource that describes what is known about the functions and diversity of plasmids in the wild? Prokaryotes or eukaryotes?
Wikipedia points to five kinds of plasmids (fertility (F),…
shigeta
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5
votes
1 answer
What volume of elution buffer for plasmid-containing filter paper?
I am attempting to elute a 9kb plasmid from a square of filter paper sent to me to be transformed into DH5a E. coli (photo below). Each filter paper square has been spotted with 1ug of plasmid DNA.
I have two questions:
AddGene's protocol states…
doremi
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3
votes
1 answer
How do you read the gene insert domains on a plasmid from Addgene?
This is a pretty basic question but I cannot find an answer anywhere. For plasmids on Addgene such as this one, there is supposed to be a GtCCR1 rhodopsin domain insert with a EYFP tag. However on the plasmid viewer shown below, there is only an…
Michael
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2
votes
1 answer
Large plasmid ligation: do I need alkaline phosphatase?
I have a doubt about a protocol I'm using for a ligation of a very large plasmid (15kbp circa) with an insert: after digestion with a single-cut enzyme (sticky ends), the protocol passes directly to the purification of the plasmid without making any…
J.G.
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2
votes
0 answers
Can we modify plasmids to eliminate antibiotic resistance?
This is a very general question regarding the potential use of artificially modified plasmids to block or modify bacterial evolution in order to prevent development of anti-biotic resistance.
For example the most crude example might be to modify a…
2
votes
2 answers
Plasmid: Empty Backbone?
When searching for a plasmid on Addgene what does it mean if it is said to be an "empty backbone"? Why would one want an empty backbone?
Kelly Eckartt
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2
votes
1 answer
The pBR322 convention
What is the pBR322 convention? For example in the description of the pET28a vector we can find information that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map.
Ala
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2
votes
1 answer
Can introns be removed when designing protein expressing episomal plasmids?
If your goal is to cause a specific protein to be expressed transiently via an episomally maintained gene, could you just exclude all introns and the exons not associated with that transcript variant when choosing what DNA to package into your…
Gumpf
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1
vote
0 answers
What is the origin of plasmids?
As I understand it, plasmids, like mitochondria, have their own genetic material and are capable of self-replication.
According to Wikipedia: Plasmids are considered replicons, units of DNA capable of replicating autonomously within a suitable…
João Maldonado
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1
vote
1 answer
Plasmid addiction system in high-copy number plasmids
Are there high-copy number plasmids that utilize a plasmid-addiction system?
kane9530
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1
vote
2 answers
Can I use TBE to elute a plasmid shipped on filter paper?
I'm receiving a plasmid shipped to me dried on sterile filter paper, as described in this question. This protocol and many others like it call for eluting the DNA from the paper using TE buffer.
My question is can I also use TBE to elute, as I…
doremi
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