Blood & Cell Culture DNA Kit

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.

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.

• 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

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.

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.).

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).

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).

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).