Blood DNA Mini Kit(<1ml)

Description

Blood genomic DNA purification can be complicated by cellular proteins, RNA, ions and other components present in whole blood. An effective whole blood DNA extraction workflow must provide sufficient sample lysis while limiting DNA degradation and removing impurities before downstream molecular biology experiments.

The Blood DNA Mini Kit is a silica membrane spin-column kit for genomic DNA extraction and purification from anticoagulated whole blood. The workflow combines Foregene Protease Plus with Buffer BL1 and Buffer BL2 for blood sample digestion, followed by selective DNA binding, washing and elution using the supplied DNA-Only Column.

The kit can process up to 1 mL of anticoagulated blood with non-nucleated red blood cells per preparation. Purified genomic DNA can be obtained in approximately 40 minutes and used for PCR, enzyme digestion, Southern hybridization and library construction.

For Research Use Only.

Key Selection Information

• Sample: Anticoagulated whole blood

• Sample input: Up to 1 mL for blood with non-nucleated red blood cells

• Purification method: Silica membrane spin column

• Processing time: Approximately 40 minutes

• Kit size: 50 preparations

• Downstream applications: PCR, enzyme digestion, Southern hybridization and library construction

Specifications

Features

• Up to 1 mL whole-blood input: Processes up to 1 mL of anticoagulated blood with non-nucleated red blood cells per preparation.

• Approximately 40-minute workflow: Provides a defined lysis, binding, washing and elution procedure for whole blood genomic DNA purification.

• Silica membrane spin-column format: The DNA-Only Column selectively binds DNA while helping remove RNA, proteins, ions and other cellular impurities.

• No additional RNase required: The protocol instructs users not to add RNase to any supplied buffer.

• No organic extraction: The procedure does not require phenol-chloroform extraction.

• No DNA precipitation step: Genomic DNA is purified through column binding, washing and elution rather than alcohol precipitation.

• Room-temperature centrifugation: Centrifugation steps are carried out at 15–25°C.

• Multiple molecular biology applications: Purified genomic DNA can be used for PCR, enzyme digestion, Southern hybridization and library construction.

Sample Compatibility

Blood with Non-Nucleated Red Blood Cells

The kit can process up to 1 mL of anticoagulated blood with non-nucleated red blood cells per preparation.

• For samples below 200 µL, Buffer BL1 is added to bring the total volume to 300 µL before proceeding with lysis.

• For samples from 200 to 1,000 µL, Buffer DC is used before the BL1 lysis step.

• Samples from 500 to 1,000 µL may be divided evenly between two tubes during Buffer DC treatment.

Blood with Nucleated Red Blood Cells

The supplied protocol specifies using 5–20 µL of anticoagulated blood containing nucleated red blood cells with 300 µL Buffer BL1.

Anticoagulants

EDTA is recommended as the blood anticoagulant. For heparin-anticoagulated blood, the manual instructs users to apply the specified Buffer DC pretreatment before continuing to the lysis step.

Applications

The Blood DNA Mini Kit is intended for genomic DNA extraction from anticoagulated whole blood for research workflows requiring purified DNA. The silica membrane spin-column procedure removes RNA, proteins, ions and other cellular impurities before DNA elution.

PCR

Purified genomic DNA can be used as template DNA for PCR-based molecular biology experiments.

Restriction Enzyme Digestion

The extracted genomic DNA can be used in enzyme digestion workflows requiring purified DNA.

Southern Hybridization

Genomic DNA purified from anticoagulated whole blood can be used for Southern hybridization experiments.

Library Construction

The purified DNA can be used as starting material for molecular biology library construction workflows.

Kit Components

*Buffer BL2 and Buffer PW contain irritating liquid salt. Wear gloves and take appropriate protective measures while handling these reagents.

Blood Genomic DNA Extraction Workflow

1. Prepare the anticoagulated whole blood sample according to its volume and red blood cell type.

2. Digest the prepared sample with Foregene Protease Plus and Buffer BL1 at 65°C.

3. Add Buffer BL2 and continue incubation at 65°C with intermittent vortex mixing.

4. Add anhydrous ethanol and mix thoroughly.

5. Transfer the complete mixture, including any flocculent precipitate, to the DNA-Only Column.

6. Centrifuge to bind genomic DNA to the silica membrane.

7. Wash the column with Buffer PW and Buffer WB1.

8. Perform an empty-column centrifugation to remove residual wash buffer.

9. Elute genomic DNA with Buffer EB preheated to 65°C.

Before You Begin

Materials and Equipment Required

• Anhydrous ethanol

• Microcentrifuge capable of 12,000 rpm, approximately 13,400 × g

• Vortex mixer

• Heating block or water bath capable of maintaining 65°C

• 1.5 mL centrifuge tubes

• 2 mL centrifuge tubes for DNA elution

Buffer WB1 Preparation

Before using catalog number DE-05111, add 60 mL of anhydrous ethanol to Buffer WB1. Confirm that ethanol has been added before beginning the DNA purification procedure.

Buffer Inspection

Inspect Buffer BL1, Buffer BL2 and Buffer PW for precipitation before use. If precipitation is present, warm the affected solution at 37°C for approximately 10 minutes and mix thoroughly until the precipitate dissolves.

Sample Handling

Avoid repeated freeze-thaw cycles. Repeated freezing and thawing may reduce the extracted DNA amount and result in smaller DNA fragments.

Storage and Stability

Store the complete kit under dry conditions at room temperature, 15–25°C, for up to 12 months. Storage at 2–8°C can support longer storage, although the manual does not specify an extended expiration period.

Foregene Protease Plus remains active at room temperature for up to three months. Storage at 4°C is recommended for improved activity and stability. Do not store Foregene Protease Plus at −20°C.

Some kit solutions may form precipitates at low temperature. Allow the solutions to equilibrate to room temperature before use. If necessary, warm the affected solution at 37°C for approximately 10 minutes and mix thoroughly until the precipitate dissolves.

Documents

Blood DNA Mini Kit Instruction Manual 

Frequently Asked Questions