QIAprep
M13 Kits use a unique silica-gel membrane that binds single-stranded DNA in the
presence of a high concentration of chaotropic salt, and allows elution in a
small volume of low-salt buffer.The QIAprep M13 system allows preparation of up to 10 µg single-stranded phage DNA. Spin and 96-well formats are available for different throughput needs.
| Format Selection Guide | |||
| No. of samples in parallel | Format | Processing | |
| QIAprep Spin M13 Kit | 1¨C24 | Spin columns | Spin or vacuum |
| QIAprep 96 M13 Kit | 48¨C96 | 96-well plates | QIAvac 96 or BioRobot |
QIAprep
M13 Kits use a unique silica-gel membrane that binds single-stranded DNA in the
presence of a high concentration of chaotropic salt, and allows elution in a
small volume of low-salt buffer.
The QIAprep M13 procedure is based on a simple bind¨Cwash¨Celute principle (see flowchart "QIAprep M13 Procedure"). M13 Precipitation Buffer is added to the phage culture supernatant to precipitate phage particles, and the sample is applied to the QIAprep membrane. Intact phage particles collected on the membrane are treated with M13 Lysis and Binding Buffer which simultaneously lyses the phage particles and allows single-stranded DNA to bind. Contaminants such as phage proteins pass through the membrane and are efficiently removed. After a brief wash, pure single-stranded DNA is eluted in low-salt elution buffer. Further concentration and desalting are not required.
QIAprep M13 Kits yield high-purity single-stranded DNA from up to 3 ml of phage supernatant with high reproducibility (see figure "Reproducible Yields with Different M13 Clones"). The eluted DNA is ideal for:
| QIAprep M13 Kit Specifications | ||
| QIAprep Spin M13 | QIAprep 96 M13 | |
| Capacity of column reservoir | 700 µl* | 1 ml |
| ssDNA binding capacity | 10 µg | 10 µg |
| Recovery | 90% | 85% |
| Elution volume | 100 µl | 100 µ |
* 2.2 ml with extension tubes, on vacuum manifolds only
Reproducible Yields with Different M13 Clones
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Parallel purification using QIAprep M13 technology of 24 samples from 6 different M13 clones grown in liquid culture in E. coli strain TG1. 5 µl of each of 4 M13 DNA preps from the indicated clones are shown. A: M13mp19; B: mouse CA-repeat clone A; C: ¦µX clone I; D: mouse CA-repeat clone B; E: M13mp19; F: ¦µX clone II; M: M13mp19 ssDNA marker |
Reliable Automated Fluorescent Sequencing
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Fluorescent sequencing of M13mp19 purified using QIAprep M13 technology. 100 ng template was sequenced with universal primer using dRhodamine chemistry, and the sequence was analyzed on an ABI PRISM 377XL Sequencer using a 36 cm WTR gel. |