Biology Forum Molecular Biology Restricion map.

last updated by blcr11 16 years ago
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    • #10318
      khushi
      Participant

      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?

    • #86711
      eddburton
      Participant

      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?

    • #86713
      khushi
      Participant

      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…..

    • #86751
      blcr11
      Participant

      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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