Restricion map.

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khushi
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Restricion map.

Post by khushi » Fri Oct 24, 2008 2:45 pm

I am trying to construct restriction map with following data..

E cor1--------2

Ecor 1+Asp 1.....1.6,.4

Asp 1....0.9,2.1,2.4,3.6.

I am trying to make linear map.Can anyone help?

eddburton
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Post by eddburton » Sat Oct 25, 2008 1:28 am

your data looks a bit ropey, I'm assuming that the numbers are lengths of fragments in terms of bases.

How have you got more fragments with a single Asp1 digest than your double digest?

Also shouldn't each digest total fragment lengths add up to the same number?

khushi
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Re: Restricion map.

Post by khushi » Sat Oct 25, 2008 3:03 am

Yeah if it would have been same length then it would have been easy to make map....that's what is confusing me.

Isolated gene from an EcoR I digest contains a single Asp 1 restriction site but no other restriction sites.The gene is inserted into a plasmid digested with Ecor 1 and the plasmid with the insert selected,grown up and plasmid isolated.A probe prepared by labeling the original gene is used to analyze southern blots of three different restriction digest.prepare a map of the insert region containing the gene and explain southern blot data?

Asp 1-----3.6,2.4,2.1,.9

Asp 1+Ecor 1------1.6,.4

Ecor 1....2

I tried to solve this..I need to make a linear map.....

blcr11
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Post by blcr11 » Mon Oct 27, 2008 3:34 pm

The Asp1 alone digest looks to be impossible to me. You should see only two bands. They most likely won’t be the same as the EcoRI + AspI double-digest as the band lengths will be determined by how far the AspI site is from the nearest vector-derived AspI site(s). The only bands you should see are the AspI-AspI fragments that carry either the 0.4 or 1.6 kb portion of the EcoRI band. You can’t see four bands unless this is a partial digest; you should see two bands, and you may only see 1 band if the AspI gods are against you.

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