Nucleic acid electrophoresis (DNA electrophoresis and RNA electrophoresis)

related topics .
(i)
. DNA
s
gel electrophoresiselectrophoresis is one of
central
cloning of molecules.
1.
. Agar
sugar gel is used to separate fragments greater than 200 to 1000bp, simple, fast and wide separation range, high resolution
2. Polyacrylamide gel is used to isolate 5-500bp fragments; good results, very high resolution, 1bp difference of DNA fragments can be separated, can accommodate a relatively large amount of DNA, for
nucleotide
polymorphism analysis.Basic processes of agarose gel electrophoresismaterials: electrophoretic buffer (commonly used TATE or TBE), electrophoretic class agarose, acetylene-plated ethyl ingot solution (
nucleic acid
display agent), sample buffer (to ensure that nucleic acid does not diffuse in the sampled hole and for electrophoresis time indicator system), horizontal gel electrophoresis device and glue system, DC power supply system.basic process: glue → put into the electrophoresis tank and add electrophoretic buffer → remove the comb→ mix the appropriate amount of nucleic acid sample and sample buffer and sample in the sample hole→ Electrophoresis →seed after the right time→ remove the gel for blycethyl bromide ingot staining→
gel imaging system
observation and photographic preservation results under ultraviolet light Note: Ethyl bromide ingots Carcinogenic, with
gloves different concentrations of agarose gel separation range



→ Factors that affect the rate at which DNA migrates in the gel: the size of the
1 DNA molecule
2 configuration
3 gel concentration
4 voltage
5 buffer

.


.

. Special Gel Electrophoresis
Inverted Electric Field Gel Electrophoresis (FIGE): DNA molecules with a molecular weight of 10 to 2000kb;
Clamp Uniform Electric Field Electrophoresis (CHEF Electrophoresis): DNA molecules greater than 107bp can be separated (ii) RNA Electrophoresis
Basic processes are the same as DNA electrophoresis, but it should be clear Because RNA molecules are very sensitive to the role of RNA enzymes, all solutions must be configured with DECC water, which inhibits RNA enzymes, and all instruments and devices in contact with RNA should be strictly treated to minimize the degradation of the sample by RNA enzymes;.