peqGOLD Blood RNA Kit - Peqlab

INTRODUCTIONpeqGOLD Blood RNA Kits are designed for isolation of total intracellular RNA from up to1.5 ml of fresh, whole blood treated with any common anticoagulant such as heparin,EDTA, or acid-citrate-dextrose. One ml of blood typically yields 1 – 5 µg of total RNA.The procedure completely removes contaminants and enzyme inhibitors making totalRNA isolation fast, less than 60 min. needed, convenient, and reliable. There is no needfor phenol/chloroform extractions, and time-consuming steps such as CsCl gradientultracentrifugation, and precipitation with isopropanol or LiCl, are eliminated.RNA purified using the peqGOLD Blood RNA method is ready for applications such asRT-PCR, Poly(A) + -RNA extractions, Northern Blots, Nuclease Protection Assays and in vitrotranslations.peqGOLD Blood RNA Kits are available as S- or C-line columns (Safety-Line, Order No.12-6814-xx or Classic Line, Order No. 12-6614-xx). Safety-Line columns can be closedtightly by lids to avoid cross-contamination more effectively. C-line columns possess no lid,allowing higher sample throughput and a more comfortable handling.PRINCIPLEThe peqGOLD Blood RNA Kits use the reversible binding properties of PerfectBind matrix,a new silica-based material. This is combined with the speed of mini-column spintechnology. Red blood cells are selectively lysed and white cells collected bycentrifugation. After lysis of white blood cells under denaturing conditions that inactivateRNases, total RNA is purified in the PerfectBind spin column. A specially formulated highsalt buffer system allows RNA molecules greater than 200 bases to bind to the matrix.Cellular debris and other contaminants (such as hemoglobin) are effectively washed awayand high quality RNA is finally eluted in RNase-free water.peqGOLD Blood RNA Kits are specially designed for isolation of total RNA out of 1.5 mlmaximum sample volume. The functionality of the kit is not restricted by the bindingcapacity of the PerfectBind RNA Column, but it is restricted by the maximum samplevolume. Due to the large quantity of erythrocytes and proteins in the blood, yield andquality of the received RNA are reduced clearly on use of larger sample volumes.It is also possible to use this kit for RNA isolation out of frozen blood sample, but thesample volume is then restricted to 150 µl for each column.peqGOLD Blood RNA Kit Art.-Nr.: 12-6814/12-6614PEQLAB_v0314 11