This kit provides a rapid spin column method for the isolation and purification of total RNA (including microRNA) from exfoliated cells that have been shed into the urine from the urinary tract. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other cellular components such as proteins, as well as from the contaminating species found in urine such as glucose and salts, without the use of phenol or chloroform. The purified RNA is of the highest integrity and can be used in a number of downstream applications including real-time RT-PCR, qRT-PCR, Northern blotting, NGS, and expression array assays. Multiple samples can be processed in 20 minutes.
Background
RNA biomarkers from exfoliated cells can be used as non-invasive tools for a number of diagnostic and research applications including the diagnosis and monitoring of bladder, kidney, or other urinary-tract cancers.
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| Kit Specifications | |
| Maximum Column Binding Capacity | 50 μg |
| Volume of Urine Processed | 1-50 mL |
| Maximum Input of Exfoliated Cells | 1 x 106 |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Time to Complete 10 Purifications | 20 minutes (for Exfoliated Cells)30 minutes (for Bacteria) |
| Average Yield | ~1μg RNA per 1 x 105 cells (For Exfoliated Cells – Varies due to cell density sample)~0.5μg RNA per 1 x 107 cells(For Bacteria – Varies due to cell density sample) |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 22550 (25 preps) |
|---|---|
| Buffer SK | 15 mL |
| Wash Solution A | 18 mL |
| Elution Buffer E | 6 mL |
| Micro Spin Columns | 25 |
| Collection Tubes | 25 |
| Elution Tubes (1.7 mL) | 25 |
| Product Insert | 1 |
The Genomic DNA Extraction Kit (HMW, Magnetic Beads) provides a reliable and fast process for extracting high molecular weight (HMW) genomic DNA from cells, blood, and tissues using Solid Phase Reversible Immobilization (SPRI) magnetic beads. With our proprietary magnetic beads technology, the kit eliminates the tedious centrifuge steps for columns. The kit provides a reliable and simple approach for high-quality genomic DNA isolation with fast magnetic response time and high binding capacity.
Cat.# 50014 Genomic DNA Extraction Kit for Cells (HMW, Magnetic Beads)
Cat.# 50015 Genomic DNA Extraction Kit for Blood (HMW, Magnetic Beads)
Cat.# 50016 Genomic DNA Extraction Kit for Tissues (HMW, Magnetic Beads)
The extracted HMW genomic DNA size ranges are dependent on the beads resuspension: 50-150 kb by tube tapping and 40-100 kb by tube vortexing. Purified DNA is recovered at high yield and high purity without RNA contamination. The typical purity ratios of A260/A280 are around 1.8-2.0, and A260/A230 are around 2.2-2.5. Purified HMW genomic DNA is suitable for applications such as long-read sequencing, linked-read genome assembly, long range PCR, optical mapping, and other general applications.
Features
A portion of the extracted genomic DNA samples were loaded on a PFGE gel with a DNA ladder indicated. Sample A: liver tissue; Sample B: intestine tissue; Sample C: whole blood; Sample D: cultured 293T cells.
Cultured Cell samples
Cultured cells are collected and are resuspended in a buffer and then lysed with a lysis buffer, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in the Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.
Blood samples
Whole blood is resuspended in the RBC buffer to remove RBC. The remaining leucocytes are lysed with a lysis buffer, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.
Tissue samples
Tissues are homogenized and lysed, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.
