Blood & Cell Culture DNA Midi Kit

For isolation of up to 100 µg high-molecular-weight DNA from blood and cultured cells
  • Reliable isolation of DNA up to 150 kb in size
  • No phenol or chloroform extractions
  • Convenient, parallel processing of multiple samples

Blood & Cell Culture DNA Kits provide gravity-flow, anion-exchange tips and buffers for the efficient purification of genomic DNA from a wide range of biological samples. The purified DNA is sized up to 150 kb with an average size of 50–100 kb.

For yeast and bacteria samples, the purchase of the Genomic DNA Buffer Set (cat.no. 19060) is required.
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Blood & Cell Culture DNA Midi Kit (25)
25 QIAGEN Genomic-tip 100/G, QIAGEN Protease, Buffers
 13343
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Genomic DNA Buffer Set
Buffers, including specific lysis buffers for yeast, bacteria, cells, blood, and tissue: Y1, B1, B2, C1, G2, QBT, QC, QF; for 75 mini-, 25 midi-, or 10 maxipreps
 19060
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The Blood & Cell Culture DNA Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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QIAGEN Genomic-tip procedure.|Genomic DNA of up to 150 kb.|
Samples are first lysed (tissue samples are mechanically disrupted) and proteins simultaneously denatured in the appropriate lysis buffer. QIAGEN Protease or Proteinase K is then added and after a suitable incubation period, lysates are loaded onto the QIAGEN Genomic-tip. DNA binds to the column while other cell constituents pass through. Following a wash step to remove any remaining contaminants, pure, high-molecular-weight DNA is eluted and precipitated with isopropanol.|Pulse-field gel electrophoresis of DNA (2 µg) purified using QIAGEN Genomic-tips. M: markers.
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Performance

The QIAGEN Genomic-tip procedure is very gentle and results in negligible DNA shearing. DNA purified with QIAGEN Genomic-tips is sized up to 150 kb with an average length of 50–100 kb (see figure "Genomic DNA of up to 150 kb"). The DNA is free of all contaminants such as RNA, protein, and metabolites, and has A260/A280 ratios between 1.7 and 1.9.

The exceptionally large size of the obtained DNA makes it especially suitable for the preparation of high-quality libraries for next generation sequencing (NGS) on different platforms and is recommended by several core facilities.

Typical yields using the Blood & Cell Culture DNA Midi Kit
Sample source Yield (µg)
Whole blood (human)
5 ml 80–100
Cultured cells (HeLa)
2 x 107 80–100

Principle

QIAGEN Genomic-tips, included in Blood & Cell Culture DNA Kits, use unique QIAGEN anion-exchange technology to purify high-molecular-weight DNA from a wide range of biological samples without phenol or chloroform. Lysis buffers are optimized for different sample types and provide immediate denaturation of proteins such as nucleases, histones, and DNA-binding proteins, as well as potentially infectious viral particles. Under the pH and low-salt conditions provided by the buffer, DNA binds to the QIAGEN Resin in the column while other cell constituents such as proteins, carbohydrates, and metabolites flow through. Purified DNA is eluted in high-salt buffer. Genomic-tips operate by gravity flow, and can be left unattended without running dry. This reduces hands-on time to a minimum and makes the procedure ideal for simultaneous processing of multiple samples.  

Procedure

Samples are first lysed (tissue samples are mechanically disrupted) and proteins simultaneously denatured in the appropriate lysis buffer (see flowchart "QIAGEN Genomic-tip procedure"). QIAGEN Protease or Proteinase K is then added and after a suitable incubation period, lysates are loaded onto the QIAGEN Genomic-tip. DNA binds to the column while other cell constituents pass through. Following a wash step to remove any remaining contaminants, pure, high-molecular-weight DNA is eluted and precipitated with isopropanol. Hands-on time for the complete procedure is just 30 minutes for blood and cultured cells.

Blood & Cell Culture DNA Kits are ready-to-use kits containing all the necessary components for purification of high-molecular-weight DNA from blood and cultured cells.

Applications

DNA purified with the Blood & Cell Culture DNA Midi Kit is well suited for use in the following applications:

Sanger and next generation sequencing
RFLP analysis
Analysis of gene targeting
Screening of transgenic animals
DNA fingerprinting studies
PCR amplification
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Feature
Specifications
Applications PCR, RFLP, blotting, screening
Elution volume 0.1–2 ml
Format Genomic tip
Main sample type Blood, cells
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA
Sample amount 5 ml/2 x 10e7
Technology Anion-exchange technology
Yield 80–100 µg

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