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Agarose Gel DNA Extraction Kit
Introduction
Agrose Gel DNA Extraction Kit. Purification of DNA fragment from agrose gel within 15 minutes.
This fast and reliable kit is designed to recover DNA from agarose gels, and purify DNA fragment from PCR, RFLP, phosphorylation, labeling and any other enzymatic reactions. DNA fragments from 200 bp to 20 kb can be purified using the ezBindTM mini column with over 85% recovery rate.
Storage and Stability
All components can be stored at room temperature. All kit components are guaranteed for 2 years from the date of purchasing.
Buffer GC contains chaotropic salts. Wear gloves and use protective eyeware when handling this solution. Buffer GC may form precipitates under cool ambient condition. Warm up the buffer at 37℃ to dissolve before use.
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Gel extraction kit
Cat#: DC3511-01 Size: 50
Purification of DNA fragment from agrose gel within 15 minutes Learn More
$ 50.00
|
Gel extraction kit
Cat#: DC3511-02 Size: 250
Purification of DNA fragment from agrose gel within 15 minutes. Learn More
$ 220.00
|
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PCR purification and gel purification protocol
Kit Contents
|
Catalog# |
DC3511-01 DC3514-01 |
DC3511-02 DC3514-02 |
|
Preps |
50 |
250 |
|
Buffer GC* |
28 mL |
130 mL |
|
DNA Wash Buffer |
12 mL |
56 mL |
|
Elution Buffer |
10 mL |
20 mL |
|
ezBind Columns |
50 |
250 |
|
Collection Tubes |
50 |
250 |
|
User Menu |
1 |
1 |
Buffer GC contains chaotropic salts. Wear gloves and use protective eyeware when handling this solution. Buffer GC may form precipitates under cool ambient condition. Warm up the buffer at 37℃ to dissolve before use.
Before Starting
Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each steps.
Important:
- R Add 48 mL (DC3514-01/DC3511-01) 100% ethanol to DNA Wash Buffer before use.
- R Add 224 mL (DC3514-02/DC3511-02) 100% ethanol to DNA Wash Buffer before use.
- R A gel slice of 100 mg equals to a volume of 100 µL.
Materials supplied by users:
- R Tabletop microcentrifuge and 1.5 mL microtubes.
- R 55-60℃ water bath (For gel extraction).
- R Vacuum manifold if use vacuum protocol.
- R 70% ethanol.
Perform all steps including centrifugation at room temperature!
PCR Purification or Gel Extraction Spin Protocol
- 1. For cycle-pure (PCR reaction): Determine the PCR product on gel. Add 2 volumes of Buffer GC to 1 volume of PCR reaction and mix completely by vortexing. Briefly spin the tube to collect any drops from the inside wall and tube lid. For PCR products less than 200 bp in length, add 5 volumes of Buffer GC.
For agarose gel: Excise the DNA fragment from the agarose gel and weigh it in a 1.5 mL microtube (A gel slice of 100 mg approximately equals to 100 µL). Add 1 volume of Buffer GC to the 1.5 mL microtube and incubate the mixture at 55-60℃ for 8 min with mixing the tube by tapping the bottom of the tube every 2 min till the gel has melted completely. Cool the tube to RT.
- 2. Transfer up to 700 µL DNA/Buffer GC mixture to a spin column with a collection tube. Centrifuge at 13,000 x g for 1 min at room temperature. Discard the flow-through and put the column back to the collection tube. Repeat this step to process the remaining solution.
- 3. Add 500 µL DNA Wash Buffer to the column and centrifuge at 13,000 x g for 1 min at room temperature. Discard the flow through.
Ensure that ethanol has been added to DNA Wash Buffer as instructed before use.
- 4. Repeat step 3.
- 5. Centrifuge the empty column with the lid open at top speed for 1-3 min to remove the residual ethanol in the column.
This is critical for optimal DNA yield.
- 6. Optional: Spin the empty column with the lid open in the collection for another 1 min at top speed.
- 7. Place the column into a clean 1.5 mL micocentrifuge tube and add 30-50 µL Elution Buffer or ddH2O to the column. Incubate at room temperature for 1 min. Centrifuge at 13,000 x g for 1 min to elute the DNA.
PCR Purification or Gel Extraction Vacuum/Spin Protocol
- 1. For cycle-pure (PCR reaction): Add 2 volumes of Buffer GC to 1 volume of the PCR reaction and mix completely by vortexing. Briefly spin the tube to collect any drops from the inside wall and tube lid. For PCR products less than 200 bp, add 5 volumes of Buffer GC to 1 volume of PCR reaction
For agarose gel: Excise the DNA fragment from the agarose gel and weigh it in a 1.5 mL microtube (A gel slice of 100 mg approximately equals to 100 µL). Add 1 volume of Buffer GC to 1 volume of gel to the 1.5 mL microtube and incubate the mixture at 55-60℃ for 8 min. Mixing the tube by tapping the bottom every 2 min till the gel has melted completely. Cool the tube to RT.
- 2. Prepare the vacuum manifold according to manufacturer’s instructions. Attach the spin column to the manifold.
- 3. Load the DNA/Buffer GC mixture to a spin column.
- 4. Turn on the vacuum to let the solution pass through the column.
- 5. Wash the column by adding 500 µL of DNA Wash Buffer.
- 6. Repeat step 5. Vacuum the column for 2 min.
- 7. Put the column in a collection tube with the lid open and spin at top speed for 2 min. It is critical to remove the residual ethanol for optimal DNA yield.
- 8. Put the column to a clean 1.5 mL tube and add 30–50 µL Elution Buffer or ddH2O. Incubate at room temperature for 1 min. Centrifuge the tube at 13,000 x g to elute DNA.
Trouble Shooting Guide
|
Problem |
Possible reason |
Suggested improvement |
|
Low DNA yield |
|
Determine the volume of Buffer GC to be used correctly as instructed. For PCR products less than 200 bp, add 2 volumes Buffer GC. Make sure to set the water bath to 55-60℃ to allow gel to melt completely. Add more buffer GC if necessary. |
|
No DNA yield |
Forgot to add ethanol to DNA Wash Buffer |
Add absolute ethanol to DNA Wash Buffer as instructed before use. |
|
DNA sample floats out of well while loading agarose gel |
Ethanol was not completely removed from the column following wash step |
After the wash step, centrifuge the empty column with the lid open at top speed for 1-3 min. Repeat once. |
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