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Enzyme inactivation

Enzyme inactivation

The disappearance of an enzymes activity during in vitro conditions, such as during a lab preparation of the enzyme, where the enzyme is exposed to conditions not normally found within its environment inside a living cell (like different ph, excess or too little salt, temperature changes, etc.)

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restriction reaction on agarose gel slices

... 3. gel extraction of insert of interest 4. restriction reaction 5. enzyme removal <- I often use KpnI/BamHI and can't go with heat inactivation 6. ligation with backbone and transformation I know that restriction can be performed ...

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by dustman
Fri May 25, 2012 2:54 pm
Forum: Molecular Biology
Topic: restriction reaction on agarose gel slices
Replies: 2
Views: 1803

Inactivate an enzyme

... your opignon about what i've done. Is it correct to don't inactivate enzyme (phosphatase) after a diphosphatasing of a plasmid and do the next step which is ligation, without inactivation ? is this could affect my results ?

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by foufoulette
Wed Apr 11, 2012 1:53 am
Forum: Molecular Biology
Topic: Inactivate an enzyme
Replies: 7
Views: 2417

clonıng problem...

... without seeing everything. Are you certain that the restriction enzymes have been inactivated as well as they can be, either by heat inactivation and/or phenol/CIAA extraction? BamH1 is sometimes a fussy enzyme; are you certain ...

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by blcr11
Thu Sep 04, 2008 3:32 pm
Forum: Molecular Biology
Topic: clonıng problem...
Replies: 3
Views: 2330

Re: problem with cloning/restriction digest

... + NO INSERT + ligase. All the ligations were treated with a third enzyme to remove any background and were NOT heat inactivated prior to transformation ( I read somewhere else that heat inactivation prior to transformation lowers the efficiency of transformation). And ...

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by ashlie23
Sat Feb 23, 2008 4:53 pm
Forum: Molecular Biology
Topic: problem with cloning/restriction digest
Replies: 35
Views: 41407

Tertiary structure + kinetic energy

... and the structure that binds with substrate. This causes activation or inactivation of enzyme............. I think I m very right here.....

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by sachin
Sun Dec 24, 2006 4:11 pm
Forum: Molecular Biology
Topic: Tertiary structure + kinetic energy
Replies: 3
Views: 3005
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