Navigation Links
Easy Cosmid DNA Purification with StrataPrep Kits

Miniprep and midiprep quantities of pure high-kb DNA

Jeff Braman Scott Basehore

The StrataPrep plasmid miniprep and midiprep kits are designed to isolate up to 20 and 350 g, respectively, of high-purity plasmid DNA from a single spin cup.1,2 These kits can also easily purify very large plasmids, including cosmid DNA.

To demonstrate cosmid purification, a SuperCos I clone containing a 40-kb insert from human chromosome 11 was isolated from two, 100-ml overnight cultures using the StrataPrep EF plasmid midiprep kit. The cosmid DNA yield from the two cultures was 45 and 51 g, respectively. The A260/A280 nm ratios were 1.8 and 1.9, and the A260/A240 nm ratios were both 1.7, indicating an absence of protein, SDS, and EDTA contaminants.3 Contaminating chromosomal DNA was also absent. The resulting cosmid DNA was digested to completion with EcoR I (Figure 1).


Cosmid DNA can be also be isolated from 1.5-ml overnight cultures using the StrataPrep miniprep kit. The cosmid DNA is completely digested with restriction enzymes such as EcoR I (Figure 2).1


The midiprep purified cosmid DNA was subjected to automated fluorescent sequencing, a very sensitive application demanding pure template DNA. A representation of the high-quality sequence is shown in Figure 3. The read length was approximately 630 bases with only four sequence ambiguities.


The StrataPrep plasmid miniprep and midiprep kits both produce high yields of all sizes of DNA from 2.9-kb plasmids to 45-kb cosmids. Both have rapid protocols using a modified alkaline lysis method to yield quality sequence with automated or manual methods. The amount of purified DNA required determines which kit to use. StrataPrep purified cosmid DNA can serve as the source material for subclone library construction used in shotgun sequencing projects.4

  1. Basehore, S. and Braman, J. (1998) Strategies 12: 5-7.
  2. Braman, J. and Basehore, S. (1997) Strategies 10: 84-86.
  3. Held, P. (1998) Pharmaceutical Laboratory, February issue.
  4. Wilson, R. K. and Mardis, E. (1997) In B. Birren, et al (Eds.), Genome Analysis, A Laboratory Manual. pp. 397-403, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.



Page: All 1 2

Related biology technology :

1. Isolation of Cosmids using Eppendorfs Perfectprep Plasmid Mini Kit
2. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
3. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
4. Yeast Protein Production System Features High Yields and One-Step Purification
5. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
6. MinElute PCR Purification Kit
7. QIAquick PCR Purification Kit
8. Microarray Target Purification Kit and the Microarray Target Synthesis Kit (T7)
9. Double-Affinity Purification of GST-His Tagged Proteins Using Vivapure 8-to-96 Well Cobalt-Chelate Kit
10. Ultra-Fast Purification of Small Nucleic Acids
11. RNAClean - RNA & cDNA in vitro Reaction Purification
Post Your Comments:

(Date:11/24/2015)... , Nov. 24, 2015 /PRNewswire/ - Aeterna Zentaris ... today that the remaining 11,000 post-share consolidation (or ... Warrants (the "Series B Warrants") subject to the ... on November 23, 2015, which will result in ... giving effect to the issuance of such shares, ...
(Date:11/24/2015)... ... November 24, 2015 , ... In harsh industrial ... points for in-line sensors can represent a weak spot where leaking process media ... of retractable sensor housings , which are designed to tolerate extreme process conditions. ...
(Date:11/24/2015)... ... November 24, 2015 , ... Creation Technologies would ... named to Deloitte's 2015 Technology Fast 500 list of the fastest growing companies ... FDA-cleared, Class II medical device that speeds up orthodontic tooth movement by as ...
(Date:11/24/2015)... , Nov. 24, 2015 Capricor ... company focused on the discovery, development and commercialization of ... Ph.D., Chief Executive Officer, is scheduled to present at ... 2015 at 10:50 a.m. EST, at The Lotte New ... . . --> ...
Breaking Biology Technology:
(Date:11/18/2015)... , November 18, 2015 ... published a new market report titled  Gesture Recognition Market ... Forecast, 2015 - 2021. According to the report, the global gesture ... is anticipated to reach US$29.1 bn by 2021, at ... North America dominated the global ...
(Date:11/17/2015)... Nov. 17, 2015 Pressure BioSciences, Inc. (OTCQB: ... development and sale of broadly enabling, pressure cycling technology ... industry, today announced it has received gross proceeds of ... Private Placement (the "Offering"), increasing the total amount raised ... more additional closings are expected in the near future. ...
(Date:11/12/2015)... , Nov. 11, 2015   Growing ... reliable analytical tools has been paving the way ... qualitative determination of discrete analytes in clinical, agricultural, ... are being predominantly used in medical applications, however, ... environmental sectors due to continuous emphasis on improving ...
Breaking Biology News(10 mins):