TempliPhi™ Large Construct Kit
SIGMA/GE25-6400-80 - Cytiva 25-6400-80, sufficient for 1000 reactions
Synonym: Phi29 DNA polymerase; RCA kit; amplification kit; circular DNA template amplification kit; isothermal rolling circular amplification; nucleic acid replication system
Product Type: Chemical
| manufacturer/tradename | Cytiva 25-6400-80 |
| shipped in | dry ice |
| storage temp. | −70°C |
| usage | sufficient for 1000 reactions |
| Analysis Note: | To view the Certificate of Analysis for this product, please visit www.cytiva.com   . |
| Application: | The TempliPhi™ Large Construct DNA Amplification Kit (TempliPhi™ LC Kit) was developed specifically to prepare templates for BAC or fosmid DNA sequencing. The TempliPhi™ method utilizes bacteriophage Phi29 DNA polymerase enzyme to exponentially amplify single- or double-stranded circular DNA templates by rolling circle amplification (RCA). This isothermal amplification method produces microgram quantities of DNA from picogram amounts of starting material in a few hours. Amplification in vitro of very small amounts of template DNA eliminates the need for overnight cell culture and conventional plasmid or M13 DNA purification. The proofreading activity of Phi29 DNA polymerase ensures high fidelity DNA replication. The starting material for amplification can be a small amount of bacterial cells containing a BAC or fosmid DNA, or any circular DNA sample. Bacterial colonies can be picked from agar plates and added directly to the TempliPhi™ LC reaction. Alternatively, microliter quantities of a saturated bacterial culture or a glycerol stock can serve as starting material. Regardless of the source of starting material, amplification is completed in 18 h at 30°C with no needed for thermal cycling. The product of the TempliPhi™ Large Construct reaction is high molecular weight, double-stranded concatemers of the circular template. The TempliPhi™ Large Construct product is double-stranded DNA that can be sequenced with forward and reverse primers. DNA amplified by the TempliPhi™ Large Construct Amplification method can be used directly in cycle sequencing reactions without any purification. |
| General description: | illustra™ TempliPhi™ Large Construct Kit. illustra™ TempliPhi™ Large Construct Kit uses isothermal rolling circle amplification (RCA) to efficiently prepare DNA sequencing templates specifically from large DNA vectors (e.g., fosmids and BACs). |
| Legal Information: | illustra is a trademark of Cytiva |
| Legal Information: | TempliPhi is a trademark of Cytiva |
| Legal Information: | TempliPhi On packaging and insert for kits: For use only as licensed by Qiagen GmbH. and Cytiva. The Phi 29 DNA polymerase may not be re-sold or used except in conjunction with the other components of this kit. See US patent numbers 5,854,033, 6,124,120, 6,143,495, 6,323,009, 5,198,543, 5,576,204, and their equivalents in other countries. On container: Subject to proprietary rights of Cytiva. May not be re-sold or used except with the other components of this kit. See US patent numbers 5,576,204, and their equivalents in other countries. |
| Preparation Note: | Please be aware this product may be shipped 90 days before the expiration date. For more information on the batch specific expiration date, please contact technical service. |
| Preparation Note: | S-re the kit at -70°C. The enzyme mix must be s-red at -70°C; all other components may be s-red at -20°C Thaw components on ice and maintain at 0°C - 4°C during handling |
| Storage Temp. | −70°C |
| UNSPSC | 41116006 |