Blood & Cell Culture DNA Kits

For isolation of up to 20 µg, 100 µg or 500 µg high-molecular-weight DNA from blood and cultured cells

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Blood & Cell Culture DNA Mini Kit (25)

Cat. No. / ID: 13323

25 QIAGEN Genomic-tip 20/G, QIAGEN Protease, Buffers

KitBuffer

Blood & Cell Culture DNA Kit

Genomic DNA Buffer Set

Column type

20/G

100/G

500/G

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Blood & Cell Culture DNA Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Reliable isolation of high-molecular-weight DNA up to 150 kb in size
No phenol or chloroform extractions
Convenient, parallel processing of multiple samples

Product Details

Blood & Cell Culture DNA Kits provide gravity-flow, anion-exchange tips and buffers for efficient isolation 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.

The purchase of the Genomic DNA Buffer Set (cat.no. 19060) is required for yeast and bacteria samples.

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 preparing high-quality libraries for next-generation sequencing (NGS) on different platforms and is recommended by several core facilities.

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. At the same time, other cell constituents such as proteins, carbohydrates and metabolites flow through. Purified DNA is eluted in a 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 Kits is well suited for use in the following applications:

Sanger and next-generation sequencing
Long-read sequencing
RFLP analysis
Analysis of gene targeting
Screening of transgenic animals
DNA fingerprinting studies
PCR amplification

Features	Blood & Cell Culture DNA Mini Kit	Blood & Cell Culture DNA Midi Kit	Blood & Cell Culture DNA Maxi Kit
Applications	PCR, RFLP, blotting, screening	PCR, RFLP, blotting, screening	PCR, RFLP, blotting, screening
Elution volume	0.1–2 ml	0.1–2 ml	0.1–2 ml
Format	Genomic-tip	Genomic-tip	Genomic-tip
Main sample type	Blood, cells	Blood, cells	Blood, cells
Processing	Manual	Manual	Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	DNA	DNA	DNA
Sample amount	1 ml/5 x 10⁶	5 ml/2 x 10⁷	20 ml/1 x 10⁸
Technology	Anion-exchange technology	Anion-exchange technology	Anion-exchange technology
Yield	15–20 µg	80–100 µg	350–400 µg

Supporting data and figures

Genomic DNA of up to 150 kb.

Pulse-field gel electrophoresis of DNA (2 µg) purified using QIAGEN Genomic-tips. M: markers.

Resources

Publications

Lack of telomerase RNA gene hTERC expression in alternative lengthening of telomeres cells is associated with methylation of the hTERC promoter.

Hoare SF; Bryce LA; Wisman GB; Burns S; Going JJ; van der Zee AG; Keith WN;

Cancer Res; 2001; 61 (1):27-32 2001 Jan 1 PMID:11196173

GATA-3 is an important transcription factor for regulating human NKG2A gene expression.

Marusina AI; Kim DK; Lieto LD; Borrego F; Coligan JE;

J Immunol; 2005; 174 (4):2152-9 2005 Feb 15 PMID:15699146

FAQ

Buffer B1 is used in combination with lysozyme or lysostaphin and proteinase K for the efficient lysis of bacteria prior to DNA purification using QIAGEN Genomic-tips. Please note this buffer is not recommended for any purification procedures using QIAGEN’s silica-membrane-based spin columns.

Buffer B1 (Bacterial Lysis Buffer 1) consists of 50 mM Tris•Cl pH 8.0; 50 mM EDTA pH 8.0; 0.5% Tween 20; 0.5% Triton-X100.

How to prepare Buffer B1: Dissolve 18.61 g Na2EDTA•2H2O and 6.06 g Tris base in 800 ml distilled water. Add 50 ml 10% Tween 20 solution and 50 ml 10% Triton X-100 solution. Adjust the pH to 8.0 with HCl. Adjust the volume to 1 liter with distilled water.

FAQ ID -2944

Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample
Mix, and store at –20°C for at least 1 h to precipitate the DNA
Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15–20 min
Pour off the ethanol and wash the pellet twice with room-temperature 70% ethanol
Allow the DNA pellet to air-dry
resuspend the DNA in a suitable volume of sterile TE buffer or distilled water

FAQ ID -305

Yes, we have the following protocols:

Isolation of genomic DNA from mosquitoes or other insects using the QIAGEN Genomic tip (QG06).
Purification of total DNA from insects using the DNeasy Blood & Tissue Kit (DY14).

FAQ ID -904

Using Gentra Puregene Cell kit:

• Purification of archive-quality DNA from up to 30 Drosophila melanogaster using the Gentra Puregene Cell Kit (PG20)

• Purification of archive-quality DNA from 100 Drosophila melanogaster using the Gentra Puregene Cell Kit (PG21)

• Purification of archive-quality DNA from 300–700 Drosophila melanogaster using the Gentra Puregene Cell Kit (PG22)

Using the QIAGEN Genomic tips:

• Isolation of genomic DNA from flies using the QIAGEN Genomic-tip 100/G (QG05)

FAQ ID -1970

Buffer G2 is a component of QIAGEN Blood & Cell Culture DNA Kits, but is also provided with several other kits (e.g., for automation on the EZ1 Advanced instrument). Usually Buffer G2 is used in combination with Proteinase K or QIAGEN Protease for efficient lysis.

Buffer G2 (General Lysis Buffer ) consists of: 800 mM guanidine hydrochloride; 30 mM Tris•Cl, pH 8.0; 30 mM EDTA, pH 8.0; 5% Tween 20; 0.5% Triton X-100.

How to prepare Buffer G2: Dissolve 76.42 g guanidine hydrochloride, 11.17g Na2EDTA•2H2O, and 3.63 g Tris base in 600 ml distilled water. Add 250 ml 20% Tween 20 solution and 50 ml 10% Triton X-100 solution. Adjust the pH to 8.0 with NaOH. Adjust the volume to 1 liter with distilled water.

FAQ ID -2943

Buffer B2 is used in combination with Buffer B1, lysozyme or lysostaphin and Proteinase K for efficient lysis of bacteria prior to DNA purification using QIAGEN Genomic-tips. Genomic-tips are gravity-flow, anion-exchange columns. Please note this buffer is not recommended for any purification procedures using QIAGEN’s silica-membrane-based spin columns.

Buffer B2 (Bacterial Lysis Buffer 2) consists of 3 M guanidine hydrochloride, 20% Tween 20.

How to prepare Buffer B2: Dissolve 286.59 g guanidine hydrochloride in 700 ml distilled water. Add 200 ml 100% Tween 20. Adjust the volume to 1 liter with distilled water. pH does not need to be adjusted.

FAQ ID -2945

Yes, we have the following protocols:

Isolation of genomic DNA from frozen clotted whole blood using the QIAGEN Genomic-tip 100/G (QG02). TEST

You will need to prepare the required buffers according to the recipes in Appendix A of the QIAGEN Genomic DNA Handbook, or you can purchase the Genomic DNA Buffer Set containing pre-made solutions. Alternatively, the QIAGEN Blood & Cell Culture Midi Kit, containing Genomic-tips 100/G and buffers, can be used.

Purification of archive-quality DNA from clotted whole blood using Clotspin Baskets and the Gentra Puregene Blood Kit (PG03).
Purification of archive-quality DNA from clotted whole blood using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit (PG04).
Purification of DNA from clotted blood using the FlexiGene DNA Kit (FG01).

FAQ ID -900. Test

Yes, we have the following protocols:

Isolation of genomic DNA from fungi (culture and blood) using the QIAamp DNA Mini Kit (QA09).
Isolation of genomic DNA from plants and filamentous fungi using the QIAGEN Genomic-tip (QG08).

FAQ ID -911

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728

Bisulfite conversion of unmethylated cytosines into uracils with the EpiTect Bisulfite Kit works on DNA irrespective of the source organism. The DNA template needs to be of high purity for efficient conversion. We recommend to use genomic DNA extracted with our DNA isolation kits for clinical or animal and plant samples as a template for the EpiTect Bisulfite Kit.

FAQ ID -1209

Using QIAGEN Genomic-tips

Protocol for DNA cleanup using Genomic-tips can be found in the 'Special Applications' section of the QIAGEN Genomic DNA Handbook.

Using QIAamp DNA Micro kit

Up to 10ug of gDNA can be cleaned using the cleanup protocol in the QIAamp DNA Micro Handbook.

Using Gentra Puregene Reagents

Gentra Puregene Handbook has protocols for removal of proteins and RNA from purified gDNA in Appendix C and Appendix D respectively.

FAQ ID -618

Genomic DNA purified using QIAGEN Genomic-tips ranges in size from 20–150 kb, with an average length of 50–100 kb. Vortexing the lysate for about 20 seconds may reduce the size of the genomic DNA slightly to 20–130 kb, but can help to improve flow rates.

FAQ ID -142

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12