DNA Extraction from Blood
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实验概要
Isolation of genomic DNA from the following amounts of fresh or frozen, human blood treated with the anticoagulant EDTA or citrate. The purified genomic DNA is suitable for use in downstream applications including PCR, restriction enzyme digestion, and Southern blotting.
主要 试剂
ChargeSwitch® gDNA 100 µl Blood Kit
实验材料
Human blood samples [Details：Volume: 50-100 µl of human blood, Treatment: EDTA - or citrate-treated, Sample state: Fresh or frozen]
实验步骤
This section provides guidelines and instructions to isolate genomic DNA from 50-100 µl samples of human blood. Note that the protocol is optimized for efficient purification of DNA from these sample volumes.
1. Before Starting
Perform the following before beginning:
1) Prepare a Lysis Mix: For each sample, mix 1 ml of ChargeSwitch®Lysis Buffer (L12) and 10 µl of Proteinase K to prepare the Lysis Mix. If you are isolating DNA from multiple samples, you may scale up the volume of reagents used and prepare a master Lysis Mix.
2) Vortex the tube containing the ChargeSwitch®Magnetic Beads to fully resuspend and evenly distribute the beads in the storage buffer.
3) Prepare a Purification Mix: For each sample, mix 40 µl of ChargeSwitch® Magnetic Beads (fully resuspended; see above) and 200 µl of f you are isolating DNA from multiple samples, you may scale up the volume of reagents used and prepare a master Purification Mix.
2. Preparing the Lysate
Follow the procedure below to prepare a lysate from the 50-100 µl blood sample.
1) Transfer the 50-100 µl blood sample to a sterile microcentrifuge tube or a 96 x 2 ml deep well plate.
2) Add 1 ml of Lysis Mix (see above) to the sample and pipet up and down gently 5 times to mix.
Important: Use a 1 ml pipette tip set to 900 µl to mix the sample. Make sure that the tip is submerged, and pipet up and down gently to avoid forming bubbles.
3) Incubate the sample at room temperature for 10 minutes or until the sample is clear with no visible lumps.
4) Proceed to Binding DNA.
Isolation of genomic DNA from the following amounts of fresh or frozen, human blood treated with the anticoagulant EDTA or citrate. The purified genomic DNA is suitable for use in downstream applications including PCR, restriction enzyme digestion, and Southern blotting.
主要 试剂
ChargeSwitch® gDNA 100 µl Blood Kit
实验材料
Human blood samples [Details：Volume: 50-100 µl of human blood, Treatment: EDTA - or citrate-treated, Sample state: Fresh or frozen]
实验步骤
This section provides guidelines and instructions to isolate genomic DNA from 50-100 µl samples of human blood. Note that the protocol is optimized for efficient purification of DNA from these sample volumes.
1. Before Starting
Perform the following before beginning:
1) Prepare a Lysis Mix: For each sample, mix 1 ml of ChargeSwitch®Lysis Buffer (L12) and 10 µl of Proteinase K to prepare the Lysis Mix. If you are isolating DNA from multiple samples, you may scale up the volume of reagents used and prepare a master Lysis Mix.
2) Vortex the tube containing the ChargeSwitch®Magnetic Beads to fully resuspend and evenly distribute the beads in the storage buffer.
3) Prepare a Purification Mix: For each sample, mix 40 µl of ChargeSwitch® Magnetic Beads (fully resuspended; see above) and 200 µl of f you are isolating DNA from multiple samples, you may scale up the volume of reagents used and prepare a master Purification Mix.
2. Preparing the Lysate
Follow the procedure below to prepare a lysate from the 50-100 µl blood sample.
1) Transfer the 50-100 µl blood sample to a sterile microcentrifuge tube or a 96 x 2 ml deep well plate.
2) Add 1 ml of Lysis Mix (see above) to the sample and pipet up and down gently 5 times to mix.
Important: Use a 1 ml pipette tip set to 900 µl to mix the sample. Make sure that the tip is submerged, and pipet up and down gently to avoid forming bubbles.
3) Incubate the sample at room temperature for 10 minutes or until the sample is clear with no visible lumps.
4) Proceed to Binding DNA.
