InstaGene Matrix: Prepare DNA Templates for PCR With No Phenol/Chloroform Extractions and No Deproteinization Steps, Rev C
Prepare DNA from blood, cultured cells, or bacteria:
• Just incubate sample with InstaGene matrix at 56ºC for 15–30 min, then
boil for 8 min.
• InstaGene matrix absorbs cell lysis products, producing an improved
substrate for PCR amplification.
DNA Preparation From Whole Blood
Use whole blood samples that are frozen, refrigerated, or collected fresh.
1. Add 3–6 µl of whole blood to 1 ml of autoclaved double distilled water
in a 1.5 ml microfuge tube. Mix by inverting the tube several times.
2. Incubate the tube at room temperature for 15–30 min.
3. Spin at 10,000–12,000 rpm for 2–3 min.
4. Carefully remove all but 20–30 µl of the supernatant. Do not disturb
the pellet.
5. Add 200 µl of InstaGene matrix to the pellet and incubate at 56ºC
for 15–30 min.
Note: InstaGene matrix should be mixed at moderate speed on a magnetic
stirrer to maintain the matrix in suspension. The pipet tip used should
have a large bore, such as on a 1,000 µl pipet tip (Bio-Rad’s TBR tips,
catalog #223-9350 and 223-9351).
6. Vortex at high speed for 10 sec. Place the tube in a 100ºC heating
block or boiling water bath for 8 min.
7. Vortex at high speed for 10 sec. Spin at 10,000–12,000 rpm for 2–3
min.
8. Use 20 µl of the resulting supernatant per 50 µl PCR reaction. Store
the remainder at -20ºC. Repeat step 7 when reusing the InstaGene DNA preparation.
Note: It is important to store the prepared sample at -20ºC when not
in use.
DNA Preparation From Cultured Mammalian Cells
1. Pellet 200 µl of cells suspension from media in a microfuge tube. Spin
at 10,000–12,000 rpm for 1 min and remove the supernatant.
2. Resuspend cells in 1 ml of 1x PBS and centrifuge for 1 min at 10,000–12,000
rpm.
3. Resuspend cells in autoclaved water at 20–30 cells/µl.
4. Add 20 µl of this cell suspension to 200 µl of InstaGene matrix. Incubate
at 56ºC for 15–30 min.
Note: InstaGene matrix should be mixed at moderate speed on a magnetic
stirrer to maintain the matrix in suspension. The pipet tip used should
have a large bore, such as on a 1,000 µl pipet tip.
5. Follow the procedure for DNA preparation from whole blood, steps 6–8.
DNA Preparation From Bacteria
The protocol described below is for the preparation of genomic DNA or
episomal DNA from bacteria.
1. Pick an isolated bacterial colony and resuspend it in 1 ml of autoclaved
water in a microfuge tube.
2. Centrifuge for 1 min at 10,000–12,000 rpm. Remove the supernatant.
3. Follow the DNA preparation procedure for whole blood, steps 5–8.
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