 |
| Previous methods commonly used for this purpose include Inverse PCR (IPCR), ligation-mediated PCR (LM-PCR) and Randomly Primed PCR (RP-PCR).
Although these methods have been used efficiently and have proven to be effective,
they are still hampered by laborious steps such as restriction enzyme digestion,
adaptor ligation, and purification procedures. Moreover, arbitrary pimers in
RP-PCR often result in non-specific products due to non-specific annealing.
These drawbacks of existing genomic walking methods can be overcome by
employing Seegene's proprietary ACP™ (Annealing Control Primer)
Seegene developed DNA walking technology by incorporating ACP™ (Annealing Control Primer) technologie, which maximizes the specificities of short primers and target specific primers, respectively, and furnishes optimized data about unknown sequences.
|
 |
| A. |
The simplest method |
| |
This method requires NO complex process,
NO restriction enzyme digestion, NO immobilization, and NO complex PCR.
|
| B. |
The most reliable method |
| |
Only target products are able to be obtained. |
| |
|
| |
|
|
|
|
| A. |
Widen Application |
|
Kit allows various vector mediated transgene integration site identification
as well as unknown sequence identification |
| B. |
Higher Success Ratio |
|
Kit is highly optimized for unknown sequence identification |
| C. |
Longer PCR Product |
|
Kit generates long PCR products that you can easily analyze |
|
 |
- Confirmation of deletion or insertion
- Splicing analysis
- Confirmation of Transgene location
- Detection of proviral DNA integration sites |
|
- Cloning and sequencing of promoter region from cDNA
- Gap filling in genomic maps
- Identification of end regions of BAC or genomic DNA clone |
 |
| Methods |
DNA digestion / ligation |
Non-specific product |
Limitation |
Sensitivity |
| DNA Walking Kit |
No |
No |
No |
High |
PCRGenomeWalker
Kit (C. company) |
Yes |
From single adaptor
primer amplification |
Restriction site
Adaptor ligation |
Medium |
Vectorette or
Splinkerette PCR
(S. company) |
Yes |
PCRGenomeWalker
Kit (C. company) |
Restriction site
Adaptor ligation |
Medium |
| IPCR (Inverse PCR) |
Yes |
Vectorette or
Splinkerette PCR
(S. company) |
Restriction site
Intermolecular
ligation |
Low |
|
 |
| Product |
Cat. No. |
Size |
| DNA Walking SpeedUp™ Kit I |
K1501 |
10 rxns |
| DNA Walking SpeedUp™ Kit I |
K1502 |
25 rxns |
| DNA Walking SpeedUp™ Kit II |
K1503 |
15 rxns |
|
