Solution 1/3

A. Digest vector pQE-60 with BglII [A^GATCT] (127) and HindIII [A^AGCTT] (152).

5'--A.TCC.A/GA.TC T.CAT.CAC.CAT.CA-.---.CAC.TA/A.GCT. TAA.TTA.GCT.GAG--3'
3'--T.AGG.T CT.AG/A.GTA.GTG.GTA.GT-.---.GTG.AT T.CGA./ATT.AAT.CGA.CTC--5'

Resulting fragments following this pQE-60 digestion:

1. 3406 bp HindIII 152 - BglII 127
2.   25 bp BglII 127 - HindIII 152

B. Digest insert pGEM-GOI with BamHI [G^GATCC] (2915) and HindIII [A^AGCTT](3266).

5'--C.TCC.G/GA.TC C.ATG.GTT.CTT.CC-.---.TAG.A/AG.CT T.ATC.GAT.TTC.--3'
3'--G.AGG.C CT.AG/G.TAC.CAA.GAA.GG-.---.ATC.T TC.GA/A.TAG.CTA.AAG.--5'

Resulting fragments following this pGEM-GOI digestion:

1. 2991 bp HindIII 3266 - BamHI 2915
2.  351 bp BamHI 2915 - HindIII 3266

Let's examine this solution in more detail. Look at the table below:


pGEM-GOI Insert
pQE60 Vector

The resulting plasmid will have the following features (choose the right answer):
 
Solution number
pQE60-GOI will contain
pQE60-GOI will NOT contain
1
MCS with HindIII & NcoI
 EcoRI
2
MCS with HindIII & BamHI
 XhoI
3
 MCS with HindIII & NcoI
BglII 
4
 Two sites of NcoI
 Poly His