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Abstract for PolyATtract® System 1000
The PolyATtract® System 1000 isolates messenger RNA directly from
crude cell or tissue lysates, eliminating the need for total RNA isolations.
This system utilizes Promega's MagneSphere® technology for the
purification of 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
x 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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