What is a Qiagen Kit?
QIAGEN Plasmid Kits provide gravity-flow, anion-exchange tips for purification of transfection-grade plasmid DNA. Lysate clearing and isopropanol precipitation are achieved by centrifugation. The QIAGEN Plasmid Mega Kit (cat. 12191) can be used with the QIAfilter Mega-Giga Cartridges (cat.
How long does a Midiprep take?
Which plasmid DNA midiprep kit is right for you?
| Quick and cost-effective | Eliminates the use of chaotropic salts | |
|---|---|---|
| Purity grade | Molecular | Transfection |
| Total protocol time | 60 min | 120 min |
| Yield | Up to 200 µg | Up to 200 µg |
| Technology | Silica membrane | Anion-exchange gravity-flow column |
What happens to the chromosomal DNA in the Qiagen kit isolation procedure?
Vigorous treatment during the lysis procedure will shear the bacterial chromosome, leaving free chromosomal DNA fragments in the supernatant. The precipitated debris is removed by centrifugation or by use of a QIAfilter Cartridge, producing a cleared lysate for loading onto the QIAGEN-tip.
How do you store plasmids after miniprep?
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.
Does PCR remove primer dimers?
Primer-dimers present in PCR reactions are effectively removed during the purification process with Norgen’s PCR Purification Kit.
How do you concentrate DNA after MidiPrep?
FAQ
- Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample.
- Mix, and store at -20°C for at least 1 hour to precipitate the DNA.
- Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15-20 minutes.
How much DNA does a miniprep have?
The average DNA concentration obtained with the DirectPrep 96 Miniprep Kit is 66 ng/ul (typically >50 ng/ul) in an eluate volume of 50 ul. Up to 4 ug DNA total can be purified from 1.3 ml of LB culture medium. Please note that the actual yield will depend on the vector and host strain being used.
How long can plasmids stay room temperature?
There will be no problem to store at RT for 1 month. During mini-prep, collect your sample on nuclease-free tubes and work in a sterile environment. However, if you want to be sure, you can store it at -20C for 5 years.
What are microcentrifuge tubes for QIAGEN plasmid mini kit?
For QIAGEN Plasmid Mini Kit protocol: Microcentrifuge 1.5 ml or 2 ml microcentrifuge tubes For QIAGEN Plasmid Midi, Maxi, Mega, and Giga Kit protocols: Centrifugation tubes or vessels with suitable capacity for the volumes specified in the appropriate protocol. g with rotor for the appropriate centrifuge tubes or bottles
What is the elution buffer for QIAGEN plasmid kits?
Buffer QF is the elution buffer used in QIAGEN Plasmid Kits for plasmid purification and in QIAGEN Blood & Cell culture kits. To make 1 liter of solution, dissolve 73.05 g NaCl and 6.06 g Tris base in 800 ml distilled water. Adjust the pH to 8.5 with HCl.
How much endotoxin free DNA can you use in QIAGEN tip 100?
This protocol is for purification of up to 100 µg endotoxin-free plasmid DNA using QIAGEN-tip 100. This protocol is designed to provide up to 150 μg BAC/PAC/P1 DNA or up to 400 μg cosmid DNA. Endotoxin-free DNA is essential for gene therapy research and will improve transfection into sensitive eukaryotic cells.
Are there any toxic substances in QIAGEN plasmids?
The entire QIAGEN plasmid purification system avoids the use of toxic substances such as phenol, chloroform, ethidium bromide, and CsCl, minimizing hazard both to the user and the environment.