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Principles of Gel Electrophoresis

Gel electrophoresis is a technique used to separate macromolecules like proteins and nucleic acids based on their size, charge, or structure by placing them in an electric field. There are two main types of gels used - agarose gels and polyacrylamide gels. Agarose gels have a large separation range but low resolution, useful for separating DNA fragments between 200-50,000 base pairs. Polyacrylamide gels have a smaller separation range but very high resolution, useful for separating DNA fragments differing by a single base pair or proteins. The gel material and concentration determine factors like separation range and resolution power.

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Loay Abuhashem Alshawamreh
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367 views2 pages

Principles of Gel Electrophoresis

Gel electrophoresis is a technique used to separate macromolecules like proteins and nucleic acids based on their size, charge, or structure by placing them in an electric field. There are two main types of gels used - agarose gels and polyacrylamide gels. Agarose gels have a large separation range but low resolution, useful for separating DNA fragments between 200-50,000 base pairs. Polyacrylamide gels have a smaller separation range but very high resolution, useful for separating DNA fragments differing by a single base pair or proteins. The gel material and concentration determine factors like separation range and resolution power.

Uploaded by

Loay Abuhashem Alshawamreh
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
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/ / Principles of Gel Electrophoresis

1/2 arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/gels/principles.html
Biotechnology Index Glossary
Principles of Gel Electrophoresis
Electrophoresis is a technique used to separate and sometimes purify macromolecules
- especially proteins and nucleic acids - that differ in size, charge or conformation. A
ch, i i e f he ide-ed echie i bichei ad eca big.
When charged molecules are placed in an electric field, they
migrate toward either the positive or negative pole according to
their charge. I ca ei, hich ca hae eihe a e iie
e egaie chage, ceic acid hae a cie egaie chage
iaed b hei hhae bacbe, ad igae ad he ade.
Proteins and nucleic acids are electrophoresed within a matrix or
"gel". M c, he ge i ca i he hae f a hi ab, ih
e f adig he ae. The ge i ieed ihi a
eechei bffe ha ide i ca a ce ad e e
f bffe aiai he H a a eaie ca ae.
The gel itself is composed of either agarose or polyacrylamide, each f hich hae
aibe iabe aica a:
Agarose i a acchaide eaced f eaeed. I i ica ed a
cceai f 0.5 2%. The highe he agae cceai he "iffe" he
ge. Agae ge ae eee ea eae: i i agae de
ih bffe i, e i b heaig, ad he ge. I i a -ic.
Agarose gels have a large range of separation, but relatively low
resolving power. B aig he cceai f agae, fage f DNA
f ab 200 50,000 b ca be eaaed ig adad eecheic
echie.
Polyacrylamide i a c-ied e f acaide. The
egh f he e chai i dicaed b he cceai f
acaide ed, hich i ica beee 3.5 ad 20%.
Pacaide ge ae igifica e aig eae ha agae ge.
Becae ge ihibi he eiai ce, he be ed
beee ga ae ( cide).
Acrlamide is a potent neurotoin and should be handled ith care! Wea
diabe ge he hadig i f acaide, ad a a he
eighig de. Pacaide i cideed be -ic, b
acaide ge hd a be haded ih ge de he ibe
eece f fee acaide.
Polyacrylamide gels have a rather small range of separation, but very
/ / Principles of Gel Electrophoresis
2/2 arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/gels/principles.html
high eoling poe. In he cae of DNA, polaclamide i ed fo
epaaing fagmen of le han abo 500 bp. Hoee, nde appopiae
condiion, fagmen of DNA diffeing i lengh b a ingle bae pai ae eail
eoled. In cona o agaoe, polaclamide gel ae ed eeniel fo
epaaing and chaaceiing mie of poein.
Inde of: Gel Elecophoei of DNA and RNA
Inodcion and Inde Agaoe Gel Elecophoei of DNA
La pdaed on Jana 3, 2000
Send commen ia fom o email o [email protected]

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