Full Answer
Is there any difference between miniprep and maxiprep DNA?
DNA from midi, maxiprep kits are quite clean and of transfection quality compared to miniprep DNA. perneseblue on Feb 19 2009, 03:28 AM said: Is there any difference between miniprep and maxiprep except that you get more ug plasmid with maxiprep? Is there any difference in the procedure? 2ml verses 6L cell culture.
What is a miniprep?
The extracted plasmid DNA resulting from performing a miniprep is itself often called a "miniprep". Minipreps are used in the process of molecular cloning to analyze bacterial clones. A typical plasmid DNA yield of a miniprep is 5 to 50 µg depending on the cell strain.
What is maxiprep used for?
I think maxiprep is usually to extract transfection grade plasmid (to transfect into animal cells or ?plant cells?...not sure about the latter) because those maxi prep kits (like from Qiagen) would remove endotoxins probably due to their column and washing solutions, so the plasmid would have low endotoxin levels.
What is the difference between MIDI Prep and maxi prep?
Midi Prep: If you need 50-100 µg plasmid DNA we suggest that you order the midi prep. Maxi Prep: If you need plasmid DNA up to 1mg quantities we recommend that you order the maxi prep.
What is the difference between miniprep and Midiprep?
The mini-prep or Miniprep corresponds to the smallest quantity of plasmid DNA, between 5 and 50 µg for a culture volume of a few millilitres. The midiprep corresponds to a medium quantity of plasmid DNA, between 100 and 350 µg for a culture volume of 15-25 ml.
What is MaxiPrep used for?
Maxiprep plasmids are commonly used for mammalian cell transfections, which require high purity DNA.
What is a miniprep used for?
Mini-Prep procedure is used to isolate small plasmid DNA from bacteria while limiting contaminating proteins and genomic DNA. The plasmid quality is acceptable for restriction analysis, sequencing, cloning, or other purposes, but should not be used with out additional cleanup for embryonic injections.
Can I use miniprep DNA for transfection?
Plasmid Miniprep System gives you a new option to prepare small quantities of high-quality DNA suitable for transfection, cloning, sequencing or coupled transcription/translation.
How many lbs is Maxiprep?
To expand the starter culture to a larger culture, dilute it 1/500 to 1/1000 in LB medium containing antibiotic. For example, to make a 150 mL culture for a Maxiprep, add between 150 and 300 µL of the starter culture to 150 mL of LB medium containing antibiotic and grow for 12-16 hrs with shaking.
How long does it take to do a Maxiprep?
Which plasmid DNA maxiprep kit is right for you?Quick and cost-effectiveMost widely accepted protocol for transfectionTotal protocol time60 min120 minYieldUp to 750 µgUp to 1 mgTechnologySilica membraneAnion-exchange gravity-flow columnClarification filterNoNo4 more rows
What is a miniprep kit?
The Monarch Plasmid Miniprep Kit is a rapid and reliable method for the purification of high quality plasmid DNA. Elution in as little as 30 μl provides concentrated DNA for use in downstream applications, such as restriction digests, DNA sequencing, PCR and other enzymatic manipulations.
How much does miniprep yield?
A typical plasmid DNA yield of a miniprep is 5 to 50 µg depending on the cell strain. Miniprep of a large number of plasmids can also be done conveniently on filter paper by lysing the cell and eluting the plasmid on to filter paper.
Where does DNA go after miniprep?
Storing DNA: temperature and longevity The most common solution is to keep your plasmid at -20°C or even at -80°C, in this case your preparation can be eluted in water or in your buffer of preference, and it will be stable for years.
Is supercoiled DNA better for transfection?
Transient transfections are more efficient with highly supercoiled DNA compared to linear DNA, presumably because circular DNA is not vulnerable to exonucleases, while linear DNA fragments are quickly degraded by these enzymes (McLenachan et al., 2007; von Groll et al., 2006).
Can you miniprep mammalian cells?
Yes, it is possible to isolate plasmid DNA from mammalian cells using the QIAprep Spin Miniprep kit .
Does EDTA interfere with transfection?
In this study, we evaluated the EDTA as a new transfection method and confirmed both improved transfection efficiency and stable cell viability on hESCs. These results demonstrate that usage of EDTA is safe substitute for transfection than that of enzymatic methods.
What is a midprep?
Midipreps are used to obtain what is effectively a permanent supply of a certain plasmid, either for transformation, or for latter modification. My lab also keeps its plasmids in DNA form rather than in cells.
Does miniprep remove endotoxin?
Miniprep usually doesn't remove any endoto xin and the plasmids extracted are usually used for analysis or for transformation. -lab_member-. Mini or Maxi prep amount of DNA is depend upon the silicon packed in the column and if you endotoxin removal solution then it will be more quality than normal preparation. -bochum-.
Is miniprep cheaper than maxiprep?
miniprep column are cheaper than maxiprep, so depending on what amount of plasmid you need, you will prepare mini, midi, maxiprep. -little mouse-.
Can you resuspension a DNA pellet?
Resuspension of the DNA pellet is no longer so easy. I use a plastic dropper to resuspend my DNA gently. Over dry a maxiprep DNA pellet and you will never go home.
What is mini prep DNA?
The mini prep DNA is suitable for many applications including routine molecular biology procedures such as. Ligation and tranformation. Deliverables: The concentrated DNA which is eluted in TE buffer or water as per the customer's instructions. Most of the DNA will be supercoiled.
How much DNA do you need for maxi prep?
Maxi Prep: If you need plasmid DNA up to 1mg quantities we recommend that you order the maxi prep.
Abstract
Here we compare the PureYield™ Miniprep, Midiprep and Maxiprep Systems to other plasmid purification systems. In most cases the PureYield™ Systems outperform several other systems in terms of speed of purification, as well as quality and downstream performance of plasmid DNA.
Introduction
Bacterial plasmids are common research tools for a variety of applications including cloning, transfection, cell-free transcription/translation reactions and DNA sequencing. Most plasmid purification systems lyse bacterial cells using a modified alkaline lysis method (1) followed by purification of the plasmid by binding to a column.
Methods
The PureYield™ Plasmid Miniprep System provides high-speed purification of high-quality plasmid DNA in 10 minutes or less. Plasmid DNA can be isolated directly from up to 0.6ml of bacterial culture or from pelleted cells from up to 3ml of cell culture.
Results
We compared PureYield™ Plasmid Midiprep System to the following plasmid midiprep systems: CompactPrep® Plasmid Midi Kit, QIAfilter Plasmid Midi Kit, Qiagen® Plasmid Midi Kit and HiSpeed® Plasmid Midi Kit (all Qiagen) in terms of speed, yield, purity and performance of DNA in downstream applications.
Summary
The PureYield™ family of plasmid DNA purification systems offers extremely rapid purification of plasmid DNA without sacrificing quality and purity. In terms of speed, yield, and purity, PureYield™ Systems outperform the competitor kits in the majority of these areas.
What is a miniprep procedure?
A miniprep procedure using diatomaceous earth to bind DNA during purification and washing.
What is plasmid preparation?
A plasmid preparation is a method of DNA extraction and purification for plasmid DNA. Many methods have been developed to purify plasmid DNA from bacteria. These methods invariably involve three steps: Growth of the bacterial culture. Harvesting and lysis of the bacteria. Purification of plasmid DNA.
What is the difference between a mini and a midi?
The difference between mini and midi prep is the yield of plasmid you get from it. If the yield and quality of an mini prep is enough for your work, than there is no need to scale up. If you need several minis for one plasmid to do you experiments than I would go for midi preps.
Is midi prep endotoxin free?
As Christian indicated, mini vs. midi really just refers to the scale, but midi prep, and not mini prep, comes with an endotoxin-free possibility the last time I checked. So if you care about whether you are exposing your fish to LPS etc., then maybe you should stick with the midiprep and test for endotoxin.