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Team:Harvard/Results/Lambda Red
From 2011.igem.org
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Fig.1) Gel image illustrating that zeocin has been inserted in place of rpoZ. Colonies 1 through 4 were picked off from a zeocin plate, and were thereby selected for zeocin expression. Lanes 1, 3, 5, 7 show the 650bp band that resulted from doing PCR with rpoZ_R (a primer flanking the rpoZ site) and zeocin_R (a primer internal to the zeocin cassette). Lane 9 has no band, as the control has the endogenous rpoZ and not zeocin at that locus. Lanes 2,4,6 show the larger band when using rpoZ_F and rpoZ_R (the two primers flanking the rpoZ region) to PCR out the zeocin cassette, compared to lane 10, where the same primers are used to PCR out the smaller rpoZ gene. Lane 8 shows no band as the PCR had not worked properly, but lane 7 shows that the zeocin cassette has been inserted into this colony. | Fig.1) Gel image illustrating that zeocin has been inserted in place of rpoZ. Colonies 1 through 4 were picked off from a zeocin plate, and were thereby selected for zeocin expression. Lanes 1, 3, 5, 7 show the 650bp band that resulted from doing PCR with rpoZ_R (a primer flanking the rpoZ site) and zeocin_R (a primer internal to the zeocin cassette). Lane 9 has no band, as the control has the endogenous rpoZ and not zeocin at that locus. Lanes 2,4,6 show the larger band when using rpoZ_F and rpoZ_R (the two primers flanking the rpoZ region) to PCR out the zeocin cassette, compared to lane 10, where the same primers are used to PCR out the smaller rpoZ gene. Lane 8 shows no band as the PCR had not worked properly, but lane 7 shows that the zeocin cassette has been inserted into this colony. | ||
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[[File:zeocin insert gel.gif|frameless|915px]] | [[File:zeocin insert gel.gif|frameless|915px]] | ||
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Revision as of 02:50, 26 September 2011
Fig.1) Gel image illustrating that zeocin has been inserted in place of rpoZ. Colonies 1 through 4 were picked off from a zeocin plate, and were thereby selected for zeocin expression. Lanes 1, 3, 5, 7 show the 650bp band that resulted from doing PCR with rpoZ_R (a primer flanking the rpoZ site) and zeocin_R (a primer internal to the zeocin cassette). Lane 9 has no band, as the control has the endogenous rpoZ and not zeocin at that locus. Lanes 2,4,6 show the larger band when using rpoZ_F and rpoZ_R (the two primers flanking the rpoZ region) to PCR out the zeocin cassette, compared to lane 10, where the same primers are used to PCR out the smaller rpoZ gene. Lane 8 shows no band as the PCR had not worked properly, but lane 7 shows that the zeocin cassette has been inserted into this colony.
