Centrifuge the paxgene blood rna tube for 10 min at 30005000 x g using a swingout rotor e. We stored blood samples for up to 7 days at room temperature in paxgene blood ccfdna tubes and used these plasma supernatants and plasma from immediately processed edta tubes so far the gold standard from the same cancer patients for molecular analysis of cellfree dna. Pdf isolation of microarraygrade total rna, microrna. The stabilized blood is pelleted and washed, subjected to a.
Magmax for stabilized blood tubes rna isolation kit. Relative transcript levels of fos and il1b were determined by realtime, duplex rtpcr, using 18s rrna as an internal standard. The paxgene blood rna system is the first ivd marketed product for the collection, storage, and transportation of blood and stabilization of intracellular rna in a closed tube with subsequent isolation and purification of intracellular rna from whole blood for rtpcr used in molecular diagnostic testing. Paxgene blood dna tubes are distributed by bd and its distributors paxgene blood dna tube dna yield median and range. Total rna 18 nucleotides including mirna is then purified using the paxgene blood mirna kit. Using microarrays as readout of rna from stimulated whole blood we found a common set of expressed transcripts in rna samples from either paxgene or tempus.
Rnastabilized whole blood samples but not peripheral. Rna extraction from rnastabilizing blood collection tubes. For example, the main blood rna collection systemsthe paxgene and tempus tubesdifferently influence rna expression signatures. Add 5 ml rnasefree water to the pellet, and close the tube using a fresh secondary hemogard closure. Allow at least 10 seconds for a complete blood draw to take. How to collect blood using the a blood rna tube paxgene. The paxgene blood rna tube is used to protect rna of whole blood, and the stability of rna in this tube has already been reported. For the collection of whole blood for genomic dna isolation pdf 1655kb.
For blood collection prior to genomic dna purification using the paxgene blood dna kit. Using a blood collection set c, collect blood into the paxgene blood rna tube using your institutions recommended standard procedure for venipuncture. Total rna was purified manually with the paxgene blood rna kit. Highquality rna could be extracted from blood samples stabilized with both paxgene and tempus tubes. Modified paxgene method allows for isolation of high. Rna purification options ruo fully integrated system allows manual and automated processing. When the kit is used in conjunction with the paxgene blood rna tube, the system provides purified intracellular rna from whole blood for rtpcr used in molecular diagnostic testing.
Gene expression profiling from blood is sensitive to technology choices. The paxgene blood rna tube is of a sterile, plastic, evacuated blood collection tube containing stabilization solution tetradecyl trimethylammonium oxalate and tartaric acid. See the paxgene blood rna tube product circular for information about the use of paxgene blood rna tubes. Rna was extracted with 4 manual protocols from qiagen paxgene blood mirna kit, life technologies magmax for stabilized blood tubes rna isolation kit. How to collect blood using the paxgene blood rna tube. Allow at least 10 s for the blood draw to occur and ensure that blood has stopped flowing into the tube before removing the needle from the tube. It is part of the paxgene blood rna system that integrates the key steps of whole blood collection. This simple procedure creates a lysate that can go directly into reverse transcription and realtime pcr with the sensitivity and specificity. Maximize total rna yields from blood samples the magmax for stabilized blood tubes rna isolation kit utilizes proven magnetic beadbased technology to purify maximal yields of total rna from paxgene blood rna tubes. These are available online in convenient and compact pdf format at.
If the paxgene blood rna tube is the only tube to be drawn, blood should be drawn into a discard tube prior to drawing blood into the paxgene blood rna tube so. Pdf comparison of whole blood rna preservation tubes and. C prior to use and properly labeled with specimen identification. The paxgene blood rna tube contains an additive that stabilizes the in vivo gene transcription profile by reducing in vitro rna degradation and minimizing gene induction. Rna isolation and purification steps greatly influence the results of gene expression profiling. Qiaamp mini column tube extenders 20 ml, qiagen proteinase k, carrier rna, buffers, vacconnectors and collection tubes data demonstrating effective stabilization of blood cells.
Comparison of automated magna pure 96 and manual paxgene. However, there are few reports on the quality of rna from longterm preservation of these tubes. Immediately after blood collection gently invert the paxgene tube blood rna tube 810x. Gene expression differences between paxgene and tempus. Ensure that the paxgene blood rna tube a is at room temperature 18c25c prior to use and is properly labeled with patient. An assay using branched dna has been described to quantitatively measure mrna expression from small blood volumes, but this method did not utilise the paxgene blood rna system 8. Handbook for information about the use of paxgene blood rna tubes brt.
As the paxgene blood rna and dna tubes are not graduated or marked for volume, the creation of a volume reference tube is. The paxgene blood rna tube contains an additive that stabilizes the in vivo gene transcription profile by reducing in vitro rna. If the paxgene blood rna tube is the only tube to be drawn, blood should be drawn into a discard tube prior to drawing blood into the paxgene blood rna tube so the interior volume of the blood collection set used. Quality control of rna extracted from paxgene blood rna. Isolation of microarraygrade total rna, microrna, and dna.
Bd diagnostics, preanalytical systems 1 becton drive franklin lakes, nj, 074171885 y information department. When used in conjunction with the paxgene blood rna kit, the blood rna tube provides for accurate detection and quantitation of. Sample preservation dnarna shield effectively lyses cells, inactivates nucleases and infectious agents and is ideal for safe sample storage and transport at ambient temperatures. If the paxgene blood rna tube is the only tube to be drawn, a small amount of blood should be drawn into a discard tube a prior to drawing blood into the paxgene blood rna tube. Venous blood obtained from the retroorbital sinus of c57bl6 mice under anaesthesia was collected into heparinized tubes according to a protocol approved by the glaxosmithkline institutional animal care and use committee iacuc. The incubation time was chosen according to the preanalytix product circular paxgene blood rna kit handbook, version 2 and results obtained by rainen et al. The aim of this study was to establish a common rna isolation protocol for these two systems and investigate if it could reduce the differences in gene expression between them. Blood samples from healthy donors were collected and frozen at. Stabilized bloodtoc t nucleic acid preparation kit for. Paxgenetm blood rna tube is manufactured by bd diagnostics, preanalytical systems for preanalytix. Trix is a combination of three isolation methods figure 1, starting with 2. Levels of mir16, mir181, and mir423 were higher when blood was collected in tempus tubes than paxgene tubes when rna was extracted using the respective tubespecific protocol p0. Invitrogen magmax for stabilized blood tubes rna isolation. Our biobank conducted quality control of rna extracted from the tubes after varying storage periods.
Bd paxgene blood rna tube, bd diagnostics tube paxgene rna 2. When used in conjunction with the paxgene blood rna kit, the blood rna tube provides for accurate detection and quantitation of gene transcripts. The paxgene t m blood rna tube is of a sterile, plastic, evacuated blood collection tube containing stabilization solution tetradecyl trimethylammonium oxalate and tartaric acid. The procedure is simple and can be performed using manual or automated procedures see flowcharts manual paxgene blood rna procedure and automated paxgene blood rna procedure. Otherwise, the paxgene blood rna tube should be the last tube drawn in the phlebotomy procedure. Paxgene blood rna tubes blood was drawn into paxgene blood rna tubes from multiple donors and stored at 20 and 70c. Paxgene blood dna tube is suitable for collection of whole blood for in vitro diagnostic ivd dna testing. Blood collection tubes are sterile glass or plastic tubes with a vacuum inside facilitating the draw of a predetermined volume of liquid. Paxgene t m blood rna tubes paxgenet m blood rna kit. Hold the paxgene blood rna tube vertically, below the blood donors arm during blood collection. Thaw the paxgene blood rna tubes in a wire rack at ambient temperature 18.
A turbo dnase step efficiently eliminates contaminating gdna that can interfere with downstream applications and ensures the highest purity rna possible. Human blood rna stabilization in samples collected and. The paxgene tube reagents have been used to assess inflammatory responses in vivo using 2. Successful downstream application of the paxgene blood rna. Rna extraction from plants using the rneasy plant mini kit. Prior to blood collection, the paxgene rna stabilizer solution was removed from the bd vacutainer style paxgene blood rna tubes. Combining topnotch and uncompromising quality with an affordable price, the bd paxgene blood rna tube, bd diagnostics tube paxgene rna 2. Paxgene blood dna tubes 100, 761115 from qiagen labsave. We compared protocols for rna isolation from these novel paxgene blood collection tubes preanalytix. Blood is collected and stabilized in paxgene blood rna tubes. The paxgene blood mirna system provides a complete solution for stabilization and purification of highquality total rna 18 nucleotides including mirna from human whole blood. Thaw the paxgene blood rna tubes in a wire rack at ambient temperature 1825.
The paxgene blood rna system is available for ivd use only when composed of the paxgene blood rna tube 762165 in combination with the paxgene blood rna kit 762164 or 762174. Paxgene blood mirna kit for purification of total rna, including rna. The rnadvance blood kit is a ribonucleic acid rna isolation reagent kit built on spri paramagnetic beadbased technology. Bd paxgene blood rna tube, bd diagnostics 762165 tube.
To ensure the quality and stabilization of blood specimens and minimize the degradation of blood rna, paxgene blood rna tubes and tempus tubes containing rna stabilizing reagents are recommended. Differences in rna quality and yield between paxgene a. A blood collection set must be used with the paxgene. Endpoints to characterize the samples were rna quality and yield, and the rna transcript stability of. The paxgene blood rna kit is for the purification of intracellular rna from whole blood collected in the paxgene blood rna tube. The paxgene blood rna kit is for the purification of total rna from 2. Paxgene blood rna tube product circular ocean ridge bio. Isolation of microarraygrade total rna, microrna, and dna from a single paxgene blood rna tube. Improved preanalytical process for rna isolation from whole blood. Most commonly used to collect blood samples in venipuncture, they are designed for the collection, transport, and processing of skin puncture blood. A modified method based upon the tempus system was found to provide high yield, good posthybridization array quality, low.
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