การทำ DNA ให้บริสุทธิ์ (DNA Clean-Up)
α+ Solution™ GEL/ PCR Purification Kit
รหัสสินค้า: APGCK004 / APGCK050 / APGCK100 / APGCK300
ผู้ผลิต: ALPHAGEN
ราคา: กรุณาสอบถาม
- หลักการ
- spin column (silica matrix)
- ปริมาณตัวอย่าง
- up to 300 mg of agarose gel . up to 300 mg of agarose gel
- เวลาที่ใช้
- 40 µl
Ordering information
| APGCK004 | 4 preps |
|---|---|
| APGCK050 | 50 preps |
| APGCK100 | 100 preps |
| APGCK300 | 300 preps |
Component
| APGCK 004 | APGCK 050 | APGCK 100 | APGCK 300 | |
| DF Buffer | 1.5 ml x 2 | 40 ml | 80 ml | 240 ml |
| Wash Buffer (concentrate) | 1.0 ml | 15 ml | 25 ml | 50 ml |
| Elution Buffer | 0.5 ml | 5 ml | 6 ml | 30 ml |
| DF Column | 4 pcs | 50 pcs | 100 pcs | 300 pcs |
| Collection Tube | 4 pcs | 50 pcs | 100 pcs | 300 pcs |
| User Manual | 1 | 1 | 1 | 1 |
Brief Prodecure
Specification
Specification:
- Principle : spin column (silica matrix)
- DNA Binding capacity of spin column : 20 µg
- Sample size : up to 300 mg of agarose gel . up to 300 mg of agarose gel
- DNA size : 65 bp ~ 10 kbp
- Recovery : 70% ~ 85% for Gel extraction . 90% ~ 95% for PCR clean-up
- Binding capacity : 10 ~ 20 min
- Operation time : 40 µl
Important Notes:
- Buffer provided in this kit contain irritants. Wear gloves and lab coat when handling these buffer.
- Add the required volume of ethanol (96~100%) to Wash Buffer before use.
- Centrifugation steps are done by a microcentrifuge capable of the speed at 11,000 ~1,8000 x g.
Trouble shooting
For Gel Extraction
|
Problems |
Possible reasons |
Solutions |
|
The gel slice is hard to dissolve |
Agarose gel of high percentage (> 2 %) is used |
Add 1000 µl of DF Buffer to 1 volume of the gel slice. |
|
The size of the gel slice is too large |
If the gel slice is more than 300 mg, separate it into multiple tubes. |
|
|
Low or none recovery of DNA fragment |
The column is loaded with too much agarose gel |
The maximum volume of the gel slice is 300 mg per column. |
|
Elution of DNA fragment is not efficient |
Make sure the pH of Elution Buffer or ddH2O is between 7.0- 8.5. |
|
|
Make sure that the elution solution has been completely absorbed by the membrane before centrifuge. |
||
|
The size of DNA fragment is larger than 5 Kb |
Preheat the elution solution to 60 °C before use. |
|
|
Eluted DNA contains non-specific DNA fragment |
Contaminated scalpel |
Using a new or clean scalpel. |
|
DNA fragment is denatured. |
Incubate eluted DNA at 95 °C for 2 min, then cool down slowly to reanneal denatured DNA. |
|
|
Poor perfor- mance in the downstream applications |
Salt residue remains in eluted DNA fragment |
Wash the column twice with Wash Buffer. |
|
Ethanol residue remains in eluted DNA fragment |
Do discard the flow-through after washing with Wash Buffer and centrifuge for an additional 3 min. |
For PCR Clean-Up
|
Problems |
Possible reasons |
Solutions |
|
Low or none recovery of DNA fragment |
Apply more than 100 µl of PCR product |
If the PCR product is more than 100 µl, separate it into multiple tubes. |
|
Elution of DNA fragment is not efficient |
Make sure the pH of Elution Buffer or ddH2O is between 7.0- 8.5. |
|
|
Make sure that the elution solution has been completely absorbed by the column membrane before centrifuga- tion. |
||
|
The size of DNA fragment is larger than 5 Kb |
Preheat the elution solution to 60 °C before use. |
|
|
Poor perfor- mance in the downstream applications |
Salt residue remains in eluted DNA |
Wash the column twice with Wash Buffer. |
|
Ethanol residue remains in eluted DNA |
Do discard the flow-through after washing with Wash Buffer and centrifuge for an additional 3 min. |
Documents
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