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Total RNA Purification Kit
Name:
Total RNA Purification Kit
Type:
110210
Message:
Total RNA Purification Kit provides a rapid, safe and reliable way to purify total RNA from a wide variety of cells and tissues. The purified total RNA is free from contaminants, essentially is of full length and suitable for all applications of RNA.

Features
No phenol/chloroform extraction, no use of ethidium bromide and no CsCldensity gradient ultracentrifugation.
Extracted RNA is free from contaminants, easy to be redissolved in water or Tris buffer.
RNases in cells are rapidly inactivated.
High recovery of pure total RNA.

Principle
Buffer R-A containing guanidine iso-thiocyanate, a strong protein denaturant, lyses cells and releases total RNA into the supernatant. Meanwhile, Buffer R-A inactivates cellular RNases immediately to protect RNA from degradation. Addition of Buffer R-B to homogenate results in denaturation of protein, which forms insoluble clumps with genomic DNA. Proteins are further precipitated by the unique two-phase partition formed by addition of Buffer R-C. In this two-phase system RNA remains soluble and is separated into the lower phase. RNA is selectively precipitated when second two-phase partition is performed by addition of Buffer R-D into the lower phase. However, free genomic DNA remains soluble in lower phase and is separated from RNA when RNA is precipitated by centrifugation. Trace impurity is removed by second RNA precipitation performed in two-phase partition with Buffer R-E and Buffer R-F.

Procedure
Sample is homogenized in Buffer R-A. Add Buffer R-B to precipitate proteins. After addition of Buffer R-C, solution turns in to two phases and proteins are co-precipitated with genomic DNA and pelleted on the interface of the two phases and total RNA remains soluble and enters into the lower phase. Transfer the lower phase to a new microfuge tube and add Buffer R-D to form second two-phase system. In this step RNA is selectively precipitated from the lower phase, but sheared free genomic DNA, mitochondria l DNA, viral DNA or transfected vectors remain in the lower phase and are removed. The total RNA pellet is redissolved in Buffer R-A. Add buffer R-E and Buffer R-F for additional RNA precipitation wit h two-phase partition. The RNA pellet is then dissolved in H2O or Tris buffer and can be used for all RNA applications.

Applications
After first precipitation by Buffer R-D, the purified total RNA is suitable for RNA analysis experiments as follows:

Northern, dot or slot blotting
RNA protection detection
In vitro translation
DNA microarray-based analysis
Poly A+ RNA selection

After re-precipitated by Buffer R-F, the purified total RNA can be used for any RNA applications including RT-PCR and cDNA library construction.
Standard:
Cat. No. ------------------110210-04-----------------------------110210-08
Sample size-------4X108 cells/4 g tissues------------8X108 cells/8 g tissues
 
Parallel Genomic DNA & RNA Purification Kit
Name:
Parallel Genomic DNA & RNA Purification Kit
Type:
110250
Message:
Parallel Genomic DNA & RNA Purification Kit provides a rapid way to obtain pure RNA and genomic DNA from the same sample.

Features
No phenol/chloroform extraction, no CsCl density gradient ultracentrifugation.
Isolate pure RNA and DNA from same sample.

Principle and Procedure
The total RNA is released and isolated with two-phase partition as described in Total RNA Purification Kit. Genomic DNA remaining in the lower phase and precipitated on the interface of two phases is dissolved in Buffer G-A and selectively binds to silica membrane. Impurities are removed by washing with Buffer W1 and Buffer W2. Pure DNA is eluted in small volume of water or low-salt Tris buffer.

Applications
The total RNA is suitable for Northern blotting, RNA protection detection, in vitro translation, RT-PCR, cDNA library construction, etc. The genomic DNA can be applied in Southern blotting, PCR, RAPD, AFLP or RFLP, apoptosis analysis, etc.
Standard:
Cat. No. -----------------------------------110250-25-------------------------110250-50
Sample size--------------------2.5X108 cells/2.5 g tissue----------5X108 cells/5 g tissue
 

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