IBI Scientific MINI High-Speed Plasmid kit 300 preps IB47102

FAQ

What competitor’s kit is comparable?

QIAGENS QIAprep Spin Miniprep Kit QIAGENS QuickLyse Miniprep Kit

Can QIAGEN'S 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 IBI mini plasmid kit?

Yes, in order to achieve good yield, the E.coli must be stressed with an antibiotic.

How do I increase the Yield 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.

What’s the maximum amount of sample for the Mini Plasmid column/tube?

The maximum amount of sample for the column/tube is 4 ml.

What is the primary make-up of the Elution buffer?

The primary make-up of the buffer is Tris-HCl pH 8.5. This buffer does not contain EDTA. You can also use Pure Water or TE buffer.

Can the plasmid DNA extraction kit be applied on Gram (+) Bacteria?

Our plasmid kits are primarily designed to extract plasmid DNA from Gram (-) bacteria such as E. Coli. Gram (+) bacteria has thicker cell walls so the cell lysis buffers provided in the kit do not lyse them readily. However, extraction of plasmid DNA from Gram (+) bacteria can still be achieved with additional treatment. After re-suspending the pelleted bacterial cells in the PD1 Buffer, add lysozyme to give a final concentration of 3 to 5 mg/ml. Incubate the suspension at 37° Celsius for 30-60 minutes (or for a shorter time when 5 mg/ml lysozyme is used.) This treatment weakens the cell wall of the Gram (+) bacteria. Add PD2, and follow the rest of the protocol. For certain Gram (+) bacteria with thin cell walls, such as Lactobacillus, applying a double amount of PD1, PD2, and PD3 Buffer may be enough to lyse the cells. Yet, we still recommend treating Gram (+) bacteria with lysozyme to facilitate cell lysis.

What are the difference and/or 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.

Was the RNAse A volume increased in the Mini plasmid kits?

Yes, the amount of RNAse A was doubled on 9-23-14. The concentration remained the same – 50 mg/ml. The amount of RNAse added to the 25 ml bottle of PD1 is now 100 µl. The amount RNAse added to the 65 ml bottle of PD1 is 260 µl. This is double to what it was. This is for sequencing grade plasmid DNA.

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 other buffers are the same.