하나의 construct를 이용하여 E. coli, mammalian cells, baculovirus-infected insect cells 모두에서 6xHis-tagged protein을 발현할 수 있습니다
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AllPrep DNA/RNA/Protein Mini Kit (50)
Cat. No. / ID: 80004
For 50 preps: AllPrep DNA Spin Columns, RNeasy Spin Columns, Collection Tubes, RNase-Free Reagents and Buffers
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AllPrep DNA/RNA/Protein Mini Kit은/는 분자생물학 분야에 사용하기 위한 것입니다. 이 제품은 질병의 진단, 예방, 또는 치료용이 아닙니다.
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
Features
- PCR 조건 최적화 불필요
- Insect/mammalian cells에서 발현을 통한 post-translational modifications 획득
- One construct가 효과적으로 three expression systems을 발현시킴
Product Details
The AllPrep DNA/RNA/Protein Mini Kit allows simultaneous purification of DNA, RNA, and protein from the same cell or tissue sample. The kit can be used in combination with Allprotect Tissue Reagent, which delivers immediate stabilization of DNA, RNA, and protein in tissue samples for long-term storage without freezing. Tissues can be efficiently disrupted using a TissueRuptor or TissueLyser system.
Performance
Genomic DNA and total RNA purified using the AllPrep DNA/RNA/Protein Mini Kit deliver optimal results in real-time PCR and real-time RT-PCR analyses (see figure " Reliable real-time PCR and RT-PCR analyses"). Total protein purified using the kit is suitable for SDS-PAGE and western blotting (see figure " Reliable western blotting and SDS-PAGE").
See figures
Principle
The AllPrep DNA/RNA/Protein Mini Kit standardizes sample preparation for systems biology. DNA, RNA, and protein are all prepared from the same source, eliminating the variation inherent in preparing these analytes from different samples.There is no need to split the sample into 3 prior to purification, allowing maximum recovery of DNA, RNA, and protein. In addition, no organic solvents such as phenol and acetone are required.
Procedure
Cells or tissues are first lysed and homogenized in Buffer RLT, which immediately inactivates DNases and RNases as well as proteases to ensure isolation of intact DNA, RNA, and proteins. The lysate is then passed through an AllPrep DNA spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. The column is washed and pure, ready-to-use DNA is then eluted.
Ethanol is added to the flow-through from the AllPrep DNA spin column to provide appropriate binding conditions for RNA, and the sample is then applied to an RNeasy spin column, where total RNA binds to the membrane and contaminants are efficiently washed away. High-quality RNA is then eluted in RNase-free water.
Buffer APP, a novel aqueous protein precipitation solution, is added to the flow-through of the RNeasy spin column, and the precipitated proteins are pelleted by centrifugation. Intact total proteins are redissolved in an appropriate buffer and then ready to use in downstream applications. The kit includes Buffer ALO, which is compatible with SDS-PAGE, for dissolving the protein pellet. (See flowchart " AllPrep DNA/RNA/Protein procedure".)
See figures
Applications
The AllPrep DNA/RNA/Protein Mini Kit streamlines sample preparation by enabling simultaneous purification of genomic DNA, total RNA, and total protein from the same cell or tissue sample. Total protein purified using the kit is suitable for SDS-PAGE and western blotting. Genomic DNA and total RNA purified using the AllPrep DNA/RNA/Protein Mini Kit deliver optimal results in all downstream applications including:
- Real-time PCR analysis
- Real-time RT-PCR analysis
Supporting data and figures
Reliable real-time PCR and RT-PCR analyses.
Rat tissues were stabilized in Allprotect Reagent, and DNA and RNA were purified using the AllPrep DNA/RNA/Protein Mini Kit. The CT values in real-time PCR and real-time RT-PCR analyses were similar to those achieved with tissues stabilized in liquid nitrogen.
Specifications
| Features | Specifications |
|---|---|
| ApplicationsKO | PCR, RT-PCR,real-time PCR, real-time RT-PCR, Southern-,Northern-,Western Blotting, Microarrays, Genotyping,1D and 2D gel electrophoresis |
| Main sample type | Easy-to-lyse tissue or cells |
| Elution volume | RNA: 30 - 50µl, DNA: 100 - 200µl |
| Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein | Total RNA, genomic DNA and western-grade proteins |
| Format | Spin column |
| Yield | varies, depends on sample type |
| Time per run or per prep | 60 min |
| Processing | Manual (centrifugation) |
| Technology | Silica technology, Protein precipitation |
| Sample amount | up to 30 mg tissue or 1 x 10e7 cells |
Resources
Kit Handbooks (1)
Quick-Start Protocols (1)
Brochures & Guides (1)
Supplementary Protocols (1)
Technical Information (1)
Safety Data Sheets (1)
Certificates of Analysis (1)
FAQ
Can the AllPrep DNA/RNA/Protein Mini Kit be used with fibrous or lipid tissues?
What is the composition of Buffer ALO?
Does Buffer APP in the AllPrep DNA/RNA/Protein Mini Kit contain acetone?
What effect does homogenization have on DNA yield and integrity when using AllPrep DNA/RNA Kits?
What can be done if the protein pellet does not solubilize completely using the AllPrep DNA/RNA/Protein Mini Kit?
What is the composition of Buffer APP?
Can acetone be used for precipitation of protein from Buffer RLT lysates generated with RNeasy Kits?
Can tissue homogenized in Buffer RLT Plus instead of Buffer RLT be used with the AllPrep DNA/RNA/Protein Mini Kit?
Why does AllPrep DNA/RNA/Protein Mini kit use buffer RLT, but Allprep DNA/RNA Mini and Allprep DNA/RNA Micro kits use buffer RLT Plus? Are these buffers interchangeable?
3323 - Do you have a protocol for the AllPrep DNA/RNA/Protein Mini Kit on the QIAcube?