Plasmid Restriction Maps

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    • #6420
      sims10
      Participant

      I’m having problem drawing a plasmid map with the following combination enzymes…

      BamH1: 900/1500/3000
      Hindlll: 500/1600/3300
      BamH1 + Hindlll: 300/400/500/1200/3000

      Is there a simple way to solve this?

    • #61208
      sdekivit
      Participant

      first you gave to draw the restriction fragments from the separate enzymes. Thens you can take a look on where the restriction sites for both enzymes located from the combined enzyme data.

    • #61212
      2810712
      Participant

      yeah, the way above said get to the ans.
      see the no. of frags = no. of sites
      so first and second both have 3 sites each, but the combined case has not 3+3=6 but 5, so they sahre one site…

      now the combined result can be treated as one obtained by two stepped restriction…i mean imagine 1st BamH1 gives u 3 frags…
      now u treate these 3 frags with the Hindlll, so u get the above said 5 frags ,
      so u need to cut those only-BamH1 frags into those 5 frags…
      you can form only smaller or equal length frags from them, so u get this constiant,
      here Bam1 gives us 3000,1500,900
      and the combined treate gives 300,400,500,1200,3000

      now the 3000 seems remain intact,
      while 900 frag seems get broke into 300, and 400 frags
      and the 1500 frag into the 1200 and 300 strands

      now u get the possible distances between BamH1 and Hindlll sites…
      now try to make out the only Hind result from this to narrow down the possible plasmid map possibilities…

      or

      from combined result try to make the ‘only’ results by associating diff. strands togather…this is faster,

      to get Hindlll result
      500 is intact, 400 and 1200 associate, 300+3000 associate
      so the sequence is -[B/H]500[H] 400 [B] 1200 [H] 300[B] 3000-
      [B]= BamH1 site
      [H]= HIndlll site
      [B/H]= both

      it ws nice exercise, thanks…

    • #61239
      sims10
      Participant

      ok so I tried solving it…can you check if I’m correct…

      so at the top is 5400 bp (BamH1 site)

      starting counter-clockwise…

      500 bp (Hindlll)
      900 bp (BamH1)
      2100 bp (Hindlll)
      2400 bp (BamH1

    • #61275
      2810712
      Participant

      umm…
      i think instead of that 900 term u mean 400, as otherwise the total length gets more than 5400…
      try to recheck ur answer by predicting the fragment lengths on digestion using the plasim map u suspected…

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