Size of the plasmid (vector + insert)= 3681 bp Size of the vector= 2964bp size of insert= 717 bp The restriction site fo
Posted: Tue May 17, 2022 7:02 pm
Size of the plasmid (vector + insert)= 3681 bp
Size of the vector= 2964bp
size of insert= 717 bp
The restriction site for AlwNI in the vector is at 1569
The restriction site for AlwNI in the insert is 667
Correct orientation of insert in the plasmid (lane 6) will yield two bands of similar size. what are those sizes?
Incorrect orientation of the insert in the plasmid (lane 10) will produce two bands with one band being signifincantly ligther (smaller) that the other one. what are the bands sizes?
11 12 Lane 1, 8, and 12: 1 kb ladder Lane 2: PBS SK(+) No insert; cut with Xhol Lane 3: PBS SK(+) No insert; cut with AlwNI Lane 4: PBS SK(+) No insert; cut with Xhol and Alw NI Lane 5: PBS SK(+) GFP insert; cut with Xhol correct orientation Lane 6: PBS SK(+) GFP insert; cut with AlwNT (correct orientation Lane 7: PBS SK(+) GFP insert; cut with Xhol and Alw NI correct orientation Lane 9: PBS SK(+) GFP insert; cut with Xhol incorrect orientation Lane 10: PBS SK(+) GFP insert; cut with AlwNI (incorrect orientation Lane 11: PBS SK(+) GFP insert; cut with Xhol and Alw NI incorrect orientation
MCS GFP ORF 717 bp AWI OFP TMCS seal el I Xholends amplify GFP gene by PCR (oligos have Xhot site) -Restrict plasmid and GFP fragment with Xhol Remove phosphate groups from PSK with CIP LacZ/GFP transitional fusion ligate GFP fragment into PSK plasmid: the fragment can ligate in either orientation The Alwis is at bosition 667 in the GFP Bequence AlwNI Ah SGFP CORRECT ORIENTATION SK/GIF INCORRECT ORIENTATION OR Alw 1569 All 1569
Size of the vector= 2964bp
size of insert= 717 bp
The restriction site for AlwNI in the vector is at 1569
The restriction site for AlwNI in the insert is 667
Correct orientation of insert in the plasmid (lane 6) will yield two bands of similar size. what are those sizes?
Incorrect orientation of the insert in the plasmid (lane 10) will produce two bands with one band being signifincantly ligther (smaller) that the other one. what are the bands sizes?
- Size Of The Plasmid Vector Insert 3681 Bp Size Of The Vector 2964bp Size Of Insert 717 Bp The Restriction Site Fo 1 (40.43 KiB) Viewed 108 times
- Size Of The Plasmid Vector Insert 3681 Bp Size Of The Vector 2964bp Size Of Insert 717 Bp The Restriction Site Fo 2 (35.2 KiB) Viewed 108 times
MCS GFP ORF 717 bp AWI OFP TMCS seal el I Xholends amplify GFP gene by PCR (oligos have Xhot site) -Restrict plasmid and GFP fragment with Xhol Remove phosphate groups from PSK with CIP LacZ/GFP transitional fusion ligate GFP fragment into PSK plasmid: the fragment can ligate in either orientation The Alwis is at bosition 667 in the GFP Bequence AlwNI Ah SGFP CORRECT ORIENTATION SK/GIF INCORRECT ORIENTATION OR Alw 1569 All 1569