تويت
السلام عليكم ورحمة الله
هذه أول مشــــاركة لي في الجينات فأحببت أن تكون من ما استخلصته بنفسي, هي عبارة عن خطوات تقنية southern blotting وارجوا أن تكون صائبة لأنها مختصرة جدا وبالتأكيد لا غنى لي عن أرائكم و إضافاتكم إن وجد أي خطأ....
aim:-
# To detect large deletions, insertions or large TRE.
STEPS:-
1\ DNA denatured by chemicals or heat treatment.
2\ Cleaved into a number of small fragments by Restriction enzymes.
3\ Resulting fragments are separated out on gel electrophoresis.
4\ DNA fragments are blotted onto nylon membrane known as filter.
5\ Permanently fixed in position using UV light.
6\ Use probe which labeled by addition of biotin or radioactive 32P.
7\ Denaturizing of probe to be single stranded.
8\ Addition of probe to nylon membrane (filter) where it hybridizes to complementary sequences.
9\ DNA fragments to which probe has hybridized will be identified by Autoradiography.
10\ RESULTS,,,,
a) probe hybridized to a smaller band than normally → patient gene has deletions.
b) probe hybridized to a different fragment → mutation in a restriction site that normally cleaved by RE.
هذه أول مشــــاركة لي في الجينات فأحببت أن تكون من ما استخلصته بنفسي, هي عبارة عن خطوات تقنية southern blotting وارجوا أن تكون صائبة لأنها مختصرة جدا وبالتأكيد لا غنى لي عن أرائكم و إضافاتكم إن وجد أي خطأ....
aim:-
# To detect large deletions, insertions or large TRE.
STEPS:-
1\ DNA denatured by chemicals or heat treatment.
2\ Cleaved into a number of small fragments by Restriction enzymes.
3\ Resulting fragments are separated out on gel electrophoresis.
4\ DNA fragments are blotted onto nylon membrane known as filter.
5\ Permanently fixed in position using UV light.
6\ Use probe which labeled by addition of biotin or radioactive 32P.
7\ Denaturizing of probe to be single stranded.
8\ Addition of probe to nylon membrane (filter) where it hybridizes to complementary sequences.
9\ DNA fragments to which probe has hybridized will be identified by Autoradiography.
10\ RESULTS,,,,
a) probe hybridized to a smaller band than normally → patient gene has deletions.
b) probe hybridized to a different fragment → mutation in a restriction site that normally cleaved by RE.
,,, I HOPE IT WAS TRUE AND USEFULL ,,,
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