Abstract for PolyATtract® System 1000
Part# TM228
Printed in USA. Revised 3/09.
Instructions for Use of Products Z5400
& Z5420.
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.
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