- Item Description
- Technical Information
- Sample Size: 1 – 7ml bacterial culture
- Expectant Yield: up to 50µg pure plasmid DNA
- Operation Time: 20min or less
The I-Blue Mini Plasmid Kit was designed for rapid isolation of plasmid DNA from 1-7ml of cultured bacterial cells. I-Blue Lysis Buffer (an optional color indicator) is included with the kit in order to prevent common handling errors, ensuring efficient cell lysis and neutralization.
Click on Technical Information or Protocol below for more info.
|Specifications||I-Blue Mini Plasmid Kit|
|Binding Capacity:||50 µg|
|Culture Input:||1-7 ml|
|Culture Type:||Cultured bacterial cells|
|Plasmid Size:||1-15 kb|
|Typical Yield:||up to 50 µg|
|Elution Volume:||30-100 µl|
|Operation Time:||<15 minutes|
A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed in 20 minutes. The purified plasmid DNA is ready-for-use in restriction enzyme digestion, ligation PCR, and sequencing reactions.
The quality of the I-Blue Mini Plasmid Kit is tested on a lot-to-lot basis by isolating plasmid DNA from a 4ml overnightE. coli(DH5α) culture, containing plasmid pBluescript (A600>2U/ml). Following the purification process, a yield of more than 30µg is expected and the ratio of A260/A280 is between 1.8-2.0. The purified plasmid DNA (1µg) is used in EcoRI digestion and checked by electrophoresis.
What competitor’s kit is comparable? QIAGEN QIAprep Spin Miniprep Kit OR QIAGEN QuickLyse Miniprep Kit
Can QIAGEN reagents be used with IBI columns? Yes, anything that is “lysis-based” will work.
Is the use of ampicillin or other antibiotics during the cultivation of the E.coli necessary to achieve proper yield with the I-Blue mini plasmid kit? Yes, in order to achieve good yield, the E.coli must be stressed with an antibiotic.
How would I increase my yields for this kit? 1. Increase your Lysis time 2. Pre-heat the Elution Buffer 3. Perform the Elution step twice 4. With a large/concentrated sample, end-user can add extra PD1,2,3 buffers 5. See Trouble shooting guide in protocol.
What’s the maximum amount of sample for the I-Blue Mini Plasmid column/tube? The maximum amount of sample is going to be between 1 - 7 ml.
What is the primary make-up of the Elution buffer? The primary component of the Elution buffer is Tris-HCl pH 8.5.This buffer does not contain EDTA. Pure Water or TE buffer can also be used.
What’s the purpose of the W1 Buffer vs. the Wash Buffer? W1 contains chaotropic salt and ethanol. The chaotropic salt can inhibit and remove the enzyme activity, such as Taq polymerase and endonuclease. Wash Buffer contains low concentrations of salt and will remove salts and proteins from the final sample.
What’s the difference between the Hi-Speed Mini Plasmid Kit and the I-Blue Mini Plasmid Kits? The differences are 2 X volume of RNAse A and I-Blue Lysis Buffer in the I-Blue Mini. The larger volume of RNAse A increases the yield for larger sample volumes. Hi Speed Mini allows for 1-5 ml and the I-Blue Mini allows for 1-7 ml which will yield more plasmid DNA. All the other buffers are the same.