Si-Mag™ CD1 Kit

Tissue and Cell Genomic DNA Extraction and Purification Magnetic Beads Kit

This kit is suitable for extraction and purification of genomic DNA from various tissues and cultured cells. In the unique Tissue-Cell Lysis solution, genomic DNA can be efficiently extracted and then purified by the magnetic beads, yielding high pure genomic DNA with a ratio of OD260 / 280 between 1.75 to 1.85. The recovered genomic DNA size can be up to 60 kb. The purified genomic DNA is suitable for applications of PCR, Southern blot and sequencing, etc. The kit will work with a 48 well round bottom plates if a special magnetic frame is used. The kit can also be used with a variety of automatic nucleic acid extraction instruments or workstations.

Catalog #

Si-Mag-CD1

Precautions

Wear protective gloves, clothing, eye, and face protection. Wash hands thoroughly after handling.
Avoid freeze/thaw cycles and centrifugation which could damage the beads.
Vortex beads for about 10 seconds and mix them well with DNA containing material to ensure best performance.
Vortex samples for about 10 seconds before adding.
Elute DNA from the beads completely.

Kit Components included

Si-Mag magnetic beads 20 mL;
Tissue-Cell Lysis solution 40 mL;
Wash Solution 38 mL (add 25 mL of Isopropanol before use);
Elution Buffer 20 mL.

Materials needed but not provided with the kit

80% Ethanol in water.
Si-Mag Magnet (sold separately) or other magnetic racks compatible with vials used.
Isopropanol (ACS grade)
RNAse 100 mg/mL solution to remove RNA.

Storage

Magnetic beads should be stored at 2-8°C but other kit reagents need to be stored at room temperature. Lysis solution may turn cloudy if store in th cold room and to clear it up place the bottle into a water bath at 37°C.

Protocol

Preparation of sample. Bring frozen samples to 4°C before extraction
A. Tissue (see table below) frozen in liquid nitrogen should be grinded and transferred to a clean Eppendorf tube. Add 400 uL Tissue-Cell lysis solution, Vortex for 1-3 min.
B. Cells are washed with PBS twice in a clean Eppendorf tube. Discard PBS and add 400 uL Tissue-Cell lysis solution and Vortex for 1-3 min.

DNA Extraction

Incubate at 65°C for 15 min. vortexing the tube once after every 5 min. To remove RNA, add 5 ul of RNase A (100mg/mL).
Add 200 uL of magnetic beads into the tube.
Add 300 uL of isopropanol into the tube.
Vortex tube vigorously for 2 min or until no obvious precipitate in the solution. Then incubate the tube at RT for 2 min and after that put the tube onto the Si-Mag magnet rack for 60 seconds. Make sure the beads are collected at the bottom of the tube.
Remove supernatant by holding the magnet rack upside down or by pipetting.
Wash the beads with 600 uL of wash solution.
Vortex the tube for 30 seconds to mix well and then repeat Step 6 above.
Wash the beads with 600 uL of 80% ethanol twice.
Dry the beads at 55°C for 8 min leaving the tube open. Do not over-dry the beads.
Elute the DNA from beads with 150-200 uL of elution buffer, incubate fort 5 min and then vortex at full speed for 1 min.
Remove beads by using magnet rack, pipette DNA out and transfer to a clean tube.
Store purified DNA at -20°C for long-term storage.

Example of samples and an average yield

Cells

Count: 100 to 1x10⁷

DNA Yield: 0.1-50 µg

Tissue	Type	Weight (mg)	DNA Yield
Mammalian (human, mouse, rat, etc.)	Brain	35	20 - 30
	Heart	35	20 - 30
	Liver	35	30 - 50
	Spleen	35	50 - 70
	Kidney	35	30 - 50
	Lung	35	50 - 70
	Muscle	55	5 - 15
	Hair	5-50 (pieces)	0.1- 3
Non-mammalian	Fish	55	5 - 15
	Shrimp	55	5 - 15
	Other shellfish	55	30 - 60