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Cat. No.: RDSPMB-50 (for 50T)

Cat. No.: RDSPMB-200 (for 200T)

Description

Residual DNA Sample Preparation Kit with Magnetic Beads, also known as Residual Host Cell DNA Extraction Kit or Residual DNA Sample Pre-treatment Kit, uses magnetic beads coated with a novel nucleic acid purification medium. This kit is designed for stable, efficient, and convenient extraction and purification of residual host cell DNA, such as E. coli, yeast, insect, or mammalian cells, from various biological products, including intermediates, semi-finished, and finished products. The DNA samples prepared with this kit can be used with the related DNA residue qPCR detection kits.

Principle and Procedure: Proteins in the sample are first digested with proteinase K, releasing genomic DNA which specifically binds to the magnetic beads under certain conditions. The magnetic beads and solution can be quickly and efficiently separated using an external magnetic field (e.g., a magnetic separator). Subsequently, two washing steps remove various impurities, and finally, DNA is eluted from the magnetic beads using an elution buffer. The entire process does not require phenol-chloroform extraction or alcohol precipitation and can be completed in about 20 minutes.

This kit can be used in conjunction with residual DNA detection kits, such as the E.coli Residual DNA Detection Kit (qPCR), for residual testing. The recovery test results using the E. coli DNA Control as a simulated sample are shown in the table below.

Note 1: Recovery rates may vary depending on sample concentration, volume, sample type, and experimental procedures. The data in the table is for reference only.

Note 2: According to the "Determination Method for Exogenous DNA Residue" in the Chinese Pharmacopoeia (2020 Edition, Volume III) "Third Method: Quantitative PCR," the recovery rate of spiked samples should be between 50% and 150%.

This kit maintains a high recovery rate when extracting residual DNA at levels as low as femtograms (fg), typically without the need for adding carriers such as glycogen, yeast RNA, Poly A RNA, or Oligo dT DNA. The kit is used to prepare samples from 100μl simulated test samples (containing BSA) and corresponding spiked samples (i.e., adding a certain amount of standard to the test sample), and the sample concentration is detected using the E.coli Residual DNA Detection Kit (qPCR). The recovery rate calculation results are shown in the table below.

Note: The E. coli DNA standard curve equation used to calculate sample concentration in the above table is: Y = -3.371X + 36.80, where Y is the CT value, and X is the logarithm of the concentration of the test sample. The slope and Y-intercept are automatically generated by the fluorescence quantitative PCR instrument. The formula used to calculate the spiked recovery rate in the above table is: (Concentration of spiked sample - Concentration of test sample) × Elution volume / Spiked amount. The elution volume is 100μl, and the spiked amount is 3000fg E. coli DNA.

This kit's small package can extract total DNA from 50 samples, and the medium package can extract total DNA from 200 samples.

Components

• Magnetic Beads
• Proteinase K
• Washing Solution I (Add 7ml anhydrous ethanol before first use)
• Washing Solution II (Add 24ml anhydrous ethanol before first use)
• Elution Buffer
• Glycogen (20mg/ml, DNase free)

Storage

Proteinase K and Glycogen should be stored at -20ºC, while the rest can be stored at room temperature for one year. If not used for a long time, the magnetic beads can be stored at 4ºC for extended shelf life. Proteinase K and Glycogen can be stored at room temperature (15-25ºC) and remain effective for at least one week.

Note

• Magnetic separation devices are required, and it is recommended to use VSep™ Magnetic Separators.
• Before the first use, add anhydrous ethanol to Washing Solution I and Washing Solution II as indicated in the instructions and labels, mix well, and mark the bottles.
• Magnetic beads will settle after standing. Before use, vortex or invert the tubes several times until fully mixed.
• Before magnetic separation, gently shake the centrifuge tube to fully disperse the magnetic beads before approaching the magnetic field. If the magnetic beads stick to the walls, gently shake the liquid in the tube after the beads gather to allow the beads to flow down.
• Use the recommended sample volume. If the sample volume is too large, it may cause the magnetic beads to aggregate, which will affect washing and the purity of the extracted DNA. When magnetic beads aggregate, try to disperse them during washing to improve the extraction effect. If aggregation occurs, it is advisable to reduce the sample volume in subsequent experiments.
• Certain steps in this kit require a 55ºC water bath, so please prepare in advance.
• Unless otherwise stated, each vortex should be controlled within 5-10 seconds.
• All operations with this kit are performed at room temperature, with no need for an ice bath.
• It is recommended to use filter tips when preparing the PCR system to minimize the risk of contamination leading to false positives. 
• This product is for scientific research use only by professionals. It is not intended for clinical diagnosis or treatment, for food or drug use, or to be stored in a regular residence.
• For your safety and health, please wear a lab coat and disposable gloves during operation.

Related:

• E.coli Residual DNA Detection Kit (qPCR)
• CHO Residual DNA Detection Kit (qPCR)
• Residual DNA Sample Preparation Kit with Magnetic Beads

Only for research and not intended for treatment of humans or animals

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