PolyATtract® System 1000 Technical Manual
Literature # TM228
The PolyATtract® System 1000 isolates messenger RNA directly from crude cell or tissue lysates, eliminating the need for total RNA isolations. This system uses Promega MagneSphere® technology to purify poly(A)+ RNA, eliminating the need for oligo(dT) cellulose columns.
The PolyATtract® System 1000 forgoes lengthy ethanol precipitation steps, phenol:chloroform extractions, overnight ultracentrifugation through cesium chloride gradients and lithium chloride(LiCl) precipitations. Increased mRNA yields are obtained because the losses that occur during the organic extractions and precipitations are eliminated. The isolated mRNA is suitable for all molecular biology applications, including in vitro translation, cDNA synthesis, PCR analysis, ribonuclease (RNase) protection assays, primer extension and Northern blots. The increased yield of mRNA using this system allows the detection of low copy number RNAs in relatively small amounts of material using Northern blot analysis.
The PolyATtract® System 1000 protocol can be adjusted to the amount of starting material available, thus maximizing flexibility in the number of purifications that can be obtained. Sufficient reagents are supplied to isolate mRNA from up to two grams of starting material in any combination of reaction sizes, with up to 1 gram per isolation. As little as 5mg of tissue or 1 × 106 cultured cells can be processed. All system components are guaranteed to be free of contaminating ribonucleases when used as directed and are thoroughly tested to assure optimal performance.
Summary of Changes
The following changes were made to the 8/16 revision of this document:
1. The amount and concentration of the β-Mercaptoethanol component was changed, Sections 2, 4.C and 4.D.
2. The document design was updated.
Printed in USA. Revised 8/16.