New England Biolabs, T3013L, Monarch® gDNA Blood Lysis Buffer

Related Categories Genomic DNA Extraction & Purification,, Nucleic Acid Purification Applications Nucleic Acid Purification FAQ Q: What is the composition of each buffer provided with the Monarch® Spin gDNA Extraction Kit? A: The compositions of the buffers are proprietary. However, we can share the following information: Monarch® gDNA Tissue Lysis Buffer....................Detergent-based lysis buffer Monarch® gDNA Cell Lysis Buffer........................Guanidinium hydrochloride-based lysis buffer Monarch® gDNA Blood Lysis Buffer.....................Guanidinium hydrochloride-based lysis buffer Monarch® gDNA Binding Buffer...........................Guanidinium thiocyanate-based buffer Monarch® gDNA Wash Buffer..............................Ethanol-based wash buffer Monarch® gDNA Elution Buffer............................10 mM Tris-HCl, pH 9.0, 0.1 mM EDTA Q: What size gDNA can be purified with the Monarch® Spin gDNA Extraction Kit? A: Assuming high quality input material has been used, the peak size of gDNA isolated with this kit ranges from 50-60 kb. Typical DNA Integrity Numbers (DINs) for gDNA isolated from blood, cells or tissues are around 9.0. The Monarch gDNA purification protocol is optimized for isolation of gDNA that is larger than that isolated by other commercial kits. For example, it includes elution with preheated buffer that enhances the elution of larger pieces of DNA. Some starting materials such as buccal swabs or saliva, however, contain a significant proportion of dead cells. In those cells, apoptotic mechanisms have activated nucleases that digest the gDNA to shorter fragments. The peak size of gDNA isolated from such samples is usually in the range 20-40 kb. Q: Can I omit the RNase A digestion step from the protocol? A: Yes, the Monarch gDNA purification procedure has been designed to minimize the RNA amount that is co-purified. Typical percentages of RNA that are found in gDNA Monarch eluates when the RNase A digestion step is omitted are 1% (Blood Protocol), 10% (Cell Protocol), and 1-5% (Tissue Protocol). These low amounts typically do not affect most downstream applications. If the RNase A digestion step is included in each protocol, final RNA amounts that are co-purified with the gDNA are 0-1%, amounts that are not detectable by most methods. Q: Does it matter which anticoagulant is used in blood samples? A: No. gDNA isolation from blood with the Monarch® Spin gDNA Extraction Kit works well for blood that is treated with either EDTA, citrate or heparin.