詳細(xì)說(shuō)明
Lolina A/SAddress: Sindalsvej 30 8240 Risskov Danmark
Email: Info@lolina.dk
Website: https://lolina.dk
Product Specification
Product | 2x Lolina? Ultra-Long PCR Master Mix |
Catalog No. | NaM201008-1 |
Storage conditions | Dry ice transportation. Store at -20°C with a shelf life of 1 year. |
Product description
2× Lolina? Ultra-Long PCR Master Mix contains ligand-modified thermostable Taq DNA Polymerase fused with a 3’-5 ’ exonuclease activity factor, as well as optimized buffer system. It can amplify genome fragments up to 25 kb, cDNA gene sequences up to 14 kb, and λDNA fragments up to 40 kb. The mix includes dNTPs and Mg2+ required for amplification, eliminating various losses during the addition process. Additionally, protective agents in the system maintain stable activity even after repeated freeze-thaw cycles. The PCR product has an A-tail at the 3 ’ end, allowing direct connection to T-vectors.
Components
Product Name | Product Number | Specifications |
2× Lolina? Ultra-Long PCR Master Mix | NaM201008-1-NaM03 | 1 mL |
NaM201008-1-NaM08 | 5 × 1 mL | |
NaM201008-1-NaM50 | 50 × 1 mL |
Components Number | Components Name | NaM201008-1- NaM03 | NaM201008-1-NaM08 | NaM201008-1 -NaM50 |
NaM201008-1-A | 2× Lolina? Ultra-Long PCR Master Mix | 1 mL | 5 × 1 mL | 50 × 1 mL |
NaM201008-1-B | 25 mM MgSO4 | 500 μL | 1 mL | 5 × 1 mL |
NaM201008-1-C | 10 mM dNTPs | 100 μL | 500 μL | 5 × 1 mL |
Instructions
1. Reaction System
Components | Volume (μL) |
ddH2O | to 50 μL |
2× Lolina? Ultra-Long PCR Master Mixa) | 25 μL |
Template DNAb) | The appropriate amount |
Forward primer (10 μM) | 2 μL |
Reverse primer (10 μM) | 2 μL |
[Note]:
a) Reaction Buffer: The buffer contains 2.5 mM Mg2+ and optimized dNTPs. Additionally, the kit provides extra MgSO4 and dNTPs. For fragments larger than 20kb that are difficult to amplify, you can try adding 0.5-2 mM Mg2+ and 200-500 μM dNTPs.
b) The recommended amounts of different templates are as follows:
Types of templates
Range of fragment amounts for a 50 μL reaction system
Genomic DNA or E. coli bacterial liquid 10-1000 ng
Plasmid or viral DNA 0.5-50 ng
c) For amplifying long fragments, it is recommended to use a two-step method for annealing and extension. The annealing/extension time is set at 30 sec/kb, which can amplify most fragments. For fragments larger than 20 kb, the reaction time can be adjusted accordingly but should not exceed 60 sec/kb.
Cycle step | Temp. | Time | Cycles |
Initial denaturation | 95 °C | 3 min | 1 |
Denaturation | 98 °C | 10 sec |
35 |
Annealing | 68 °C | 30 sec/kb | |
Final extension | 72 °C | 10 min | 1 |
Notes
a) For your safety and health, please wear a lab coat and disposable gloves during operation.
b) This product is for research use only!
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