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Questions tagged [minipreps]

A method to purify small amounts of bacterial plasmids from the cells to get purified DNA at the end.

2
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0answers
55 views

When doing minipreps, do “good” plasmids produce more DNA than “bad” plasmids?

I've been doing minipreps for close to 10 years using the standard qiagen kits and DH5a e. coli. I typically do 24 minipreps at a time and then identify the good plasmids by enzyme digestion, PCR, ...
2
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2answers
3k views

What are the possible reasons to get extra unrecognized band in agarose gel electrophoresis?

I ran 0.7% agarose gel electrophoresis of my plasmid DNA sample consisting of GFP vector, using NheI and HindIII restriction enzyme. I used two types of plasmids, i.e., isolated by miniprep-alkali ...
2
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1answer
162 views

Plasmid maintenance

I have obtained some plasmids used as integration vectors, this question may apply to all plasmids. I would like to have a somewhat continuos source for these plasmids, let's say that the origin of ...
10
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4answers
3k views

Why is it sometimes difficult to resuspend E. coli in P1?

It's a curiosity question. When I'm doing minipreps after pelleting the bacteria sometimes it's very easy to resuspend them in P1 (Qiagen kit), but sometimes they form a rubbery clump that is very ...
6
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2answers
22k views

What is special about the N3 buffer for Qiagen minipreps?

For those who use Qiagen miniprep columns, they may be aware that the P3 buffer is the acetic acid step to neutralize cell lysis. They may also be aware that the Qiagen P3 buffer cannot be ...
8
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2answers
396 views

What are key factors when evaluating and comparing miniprep columns?

I'm looking to comparing different protocols for minipreps for plasmid DNA purification. What factors should I be looking at? A few things come to mind: Cost Yield Time per step Replacement with ...