What is the difference between normal agarose and low melting agarose?
The main properties of these agaroses are their low melting and gelling temperatures when compared with standard agaroses. LM agaroses have lower gel strength than standard agaroses, yet they can be handled easily. LM agaroses have higher clarity (gel transparency) than gels of standard agaroses.
What is the structure of agarose gel?
Agarose is a high-molecular-weight polysaccharide extracted from the cell walls of certain marine red algae. Chemically, it is a copolymer of 1,3-linked β-d-galactose and 1,4-linked 3,6-anhydro-α-l-galactose.
What is normal melting point agarose?
LPI/CPI protocol for the Comet Assay, Alkaline Version
| 1. | Equipment and non-chemical materials |
|---|---|
| 2.7 | 1.0% normal melting point agarose (NMPA) |
| 1 g + 100 ml dd H2O, microwave to dissolve agarose, compensate the evaporated H2O, cool down to around 60°C before use. | |
| 2.8 | Neutralizing buffer – 0.4 M Tris (Sigma T6066) |
How do you make agarose gel for PCR?
Pouring a Standard 1% Agarose Gel:
- Measure 1 g of agarose.
- Mix agarose powder with 100 mL 1xTAE in a microwavable flask.
- Microwave for 1-3 min until the agarose is completely dissolved (but do not overboil the solution, as some of the buffer will evaporate and thus alter the final percentage of agarose in the gel.
Why is the structure of agarose gel important?
Agarose gel is easy to cast, has relatively fewer charged groups, and is particularly suitable for separating DNA of size range most often encountered in laboratories, which accounts for the popularity of its use.
How does agarose gel concentration affect DNA migration?
The migration rates of DNA molecules in agarose gels are also affected by the composition of the gel. The migration rate of a DNA molecule decreases as the concentration of agarose in the gel increases.
Why is low melting agarose used?
Low melting temperature allows for the recovery of undamaged nucleic acids below denaturation temperature. Low Melting Agarose is ideal for digestion by agarose enzymes, which makes it very easy to recover large DNA fragments suitable for cloning or enzymatic processing.