Allprep RNA-DNA-miRNA-protein Extract Kit

Product Number: RNK4701

Shipping and Storage 

1.All solutions should be clear. If the ambient temperature is low, the solution may form precipitates and should not be used directly. It can be heated in a 37℃ water bath for a few minutes to restore clarity.

2.Inappropriate storage at low temperatures (4℃ or -20℃) can cause solution precipitation, affecting the effectiveness of use. Therefore, transportation and storage are carried out at room temperature (15℃ -25℃).

3.To avoid volatilization, oxidation, and pH changes caused by prolonged exposure of reagents to the air, each solution should be covered tightly in a timely manner after use.

Components

Component	Storage	RNK4701 50Preps
Buffer RLT Plus	RT	50 ml
Buffer WS1	RT	12 ml
Buffer WS 2/3	RT	10 ml
RNase-free H2O	RT	10 ml
Buffer IR	RT	25 ml
Buffer WB	RT	15ml
Buffer APP	RT	60 ml
Buffer EB	RT	10 ml
Genomic DNA adsorption column DA and collection tube	RT	50
RNA adsorption column RA and collection tube	RT	50

Description

This kit is designed to rapidly extract and isolate genomic DNA, as well as total RNA and Proten, including miRNA, from the same animal cell or tissue sample. The unique Buffer RLT Plus rapidly cleaves and inactivates cellular RNA/DNA enzymes, and then cleaves the mixture of RNA/miRNA/DNA/Protein while passing through a genomic DNA adsorption column, where genomic DNA is adsorbed and miRNA/RNA/Protein penetrates and filters through. The genomic DNA on the DNA adsorption column undergoes a series of rinsing centrifugation to obtain pure genomic DNA. After adjusting the binding conditions with ethanol, the filtered miRNA/RNA selectively adsorbs onto the miRNA/RNA adsorption column in a highly dissociated salt state, and then obtains pure miRNA/RNA through a series of rapid rinsing centrifugal elution. The filtrate was selectively precipitated to obtain Protein. On the basis of phenol free and chloroform free DNA/RNA rapid extraction technology, combined with exclusive separation technology, the miRNA/RNA/genomic DNA obtained simultaneously has high purity and does not interfere with each other. The obtained miRNA/RNA does not require DNase digestion and can be directly used for experiments such as reverse transcription PCR and fluorescence quantitative PCR. Genomic DNA can also be directly used for various downstream experiments such as Southern, enzyme digestion, PCR, etc.

Features

1.No toxic reagents such as phenol and chloroform are used, and no steps such as ethanol precipitation are required.

2.Fast and simple, the separation of miRNA/RNA/genomic DNA/Protein from a single sample can generally be completed within 1 hour.

3.The exclusive adsorption column and formula ensure effective removal of genomic DNA residues. Generally, the miRNA/RNA obtained does not require DNase digestion and can be directly used for reverse transcription PCR, fluorescence quantitative PCR, and other experiments.

4.Multiple column washes ensure high purity of miRNA/RNA/genomic DNA, which can be directly used in various downstream experiments.

Application

Suitable for rapid extraction and separation of genomic DNA and total RNA and Protein containing miRNA from the same animal cell or tissue sample (such as purchasing additional miRNA adsorption columns and matching solutions, and can also separate and extract total RNA and miRNA from the same sample to enrich miRNA), without the need for DNase digestion, RNA can be directly used for reverse transcription PCR and fluorescence quantitative PCR.

Note

1.All centrifugation steps are completed at room temperature using a traditional desktop centrifuge with a rotational speed of 13000rpm, such as Eppendorf5415 ℃ or a similar centrifuge.

2.The sample processing capacity should never exceed the processing capacity of the genomic adsorption column DA and RNA adsorption column RA, otherwise it may cause DNA residue or a decrease in production. There is a significant difference in RNA/DNA among different types of tissue cells, for example, the thymus is rich in DNA content, exceeding 5mg will exceed the column processing capacity. COS cells have abundant RNA content, exceeding 3×10⁶ cells will exceed the column adsorption capacity. So when starting to explore the experimental conditions, if the DNA/RNA content of the sample is not clear, it is better to use a smaller sample processing volume, such as cells not exceeding 3-4×10⁶ and tissues not exceeding 10mg. In the future, the processing capacity will be increased or decreased based on the sample testing situation.

3.Buffer RLT Plus, Buffer IR, Buffer WS1, and Buffer WS 2/3 contain irritating compounds. When operating, wear latex gloves to avoid contact with skin, eyes, and clothing. If it gets on the skin or eyes, rinse with plenty of water or physiological saline,

4.If this reagent kit is needed for the extraction of DNA/miRNA/RNA from plant samples, especially difficult samples with abundant secondary metabolites of polysaccharides and polyphenols, please consult technical personnel as other reagents may be required.

5.If additional miRNA adsorption columns and matching solutions are purchased, the total RNA and miRNA of the same sample can also be separated and extracted to enrich miRNA. Please consult us.

6.Regarding trace residues of DNA:

Generally speaking, any total RNA extraction reagent cannot completely avoid trace residues of DNA during the extraction process (DNase digestion cannot achieve 100% residue free). Our Allprep tissue/cell miRNA/DNA/Protein extraction kit, due to its unique buffer system and genomic DNA separation and clearance technology, has cleared the vast majority of DNA and does not require DNase digestion. It can be directly used for reverse transcription PCR and fluorescence quantitative PCR. In some special cases, such as residual DNA caused by excessive DNA content or strict mRNA expression analysis requiring fluorescence quantitative PCR, we suggest that when selecting templates and primers:

6.1.Select primers that cross introns to pass through the connecting regions in mRNA, so that DNA cannot serve as a template for amplification reactions.

6.2.Select primer pairs with different sizes of amplified products on genomic DNA and cDNA.

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