Total RNA Purification Kit (Blood/Bacterial /Yeast)

Purify total RNA with the highest quality and quantity

No phenol step required
Bind & elute all RNA irrespective of size or GC content, without bias
Fast and convenient spin column protocol
Isolate from a wide variety of specimens
Ensures fast and reproducible RNA purification

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SLS’ RNA Isolation Kit isolates total RNA from cultured cells, free of any contaminating DNA. Other sample materials, such as whole blood, yeast, and bacteria require a pre-lysis treatment.

Principle

A single reagent which contains a chaotropic salt and detergent lyses the sample and simultaneously inactivates RNases. After cell debris is removed by centrifugation, the lysate is applied to the glass fiber fleece in a Spin column. Under the buffer conditions used in the procedure, nucleic acids bind to the glass fiber fleece, while contaminating substances (salts, proteins, and other cellular contaminants) do not. DNA in the preparation is digested with DNase I directly on the filter. Brief wash-and-spin steps readily remove the digested DNA fragments and other contaminating substances. Once purified, the RNA can be easily eluted in a small volume of low-salt buffer.

Quickly process multiple samples. A single sample preparation is completed in 25 minutes, and multiple samples can be prepared in 45 minutes.

Prevent RNA loss. Precipitation or solvent extraction steps are not required.

Avoid DNA contamination. Integrated DNA digestion and DNase removal eliminates interference from genomic DNA (see Figure 1).

Obtain concentrated RNA. RNA is eluted in a 50 μL volume.

Obtain high sensitivity, reproducibility, and specificity for downstream applications.

Applications

RT-PCR
Northern blotting (see Figure 2)
RACE
Primer extension
Differential display
cDNA library construction
RNase protection assays
In vitro translation?

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Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants. For RNA isolation, the binding conditions can be optimized to ensure immobilization of all the RNA.

Typical RNA yields from different samples obtained with the SLS’ RNA Isolation Kit :

Starting Material	Sample Size	Time Required	Average RNA Yield
Cultured cells (K-562)	10⁶ cells	30 min	15 μg
Whole blood, human	200 µL	50 min, including RBC lysis	Sufficient for 10 RT-PCR reactions
Yeast (S. cerevisiae )	10⁸ cells	50 min, including lyticase digest	20 μg
Gram negative bacteria (E. coli )	10⁹ cells	90 min, including lysozyme digest	50 μg
Gram positive bacteria (B. subtilis )	10⁹ cells	90 min, including lysozyme digest	35 μg

Kit Contents:

	Storage Temperature (upon receipt)
Silica Spin Columns	15-25°C
Spin Column collection tube	15-25°C
Buffer RLB	15-25°C
Buffer RW1^§ (Concentrate)	15-25°C
Buffer RW2^§ (Concentrate)	15-25°C
DNase I Incubation Buffer	15-25°C
Buffer RE	15-25°C

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Product	Size	Catlog No.
Total RNA Purification Kit (from Blood/bacteria/yeast)	25 Preps	SCMR003
Total RNA Purification Kit (from Blood/bacteria/yeast)	50 Preps	SCMR003A

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