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thanks for the reply. I am trying to clone the gene again, and hope to get the success this time, Thanks again to all for ur time and patience.
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In Topic: cloning: band at different site than desired after RE digestion
18 January 2013 - 10:56 AM
thanks Lelee,
thanks for the reply. I am trying to clone the gene again, and hope to get the success this time, Thanks again to all for ur time and patience.
thanks for the reply. I am trying to clone the gene again, and hope to get the success this time, Thanks again to all for ur time and patience.
In Topic: cloning: band at different site than desired after RE digestion
06 January 2013 - 01:52 AM
leelee, on 06 January 2013 - 12:16 AM, said:
Can you post up a gel pic of both your PCR and your digests?
It is worth considering that the unwanted bands you are seeing is uncut plasmid.
How exactly do you do your PCR from your clones? What is your template (e.g. scrape of colony in water, or mini-prep), and how much are you using for your PCR?
It is worth considering that the unwanted bands you are seeing is uncut plasmid.
How exactly do you do your PCR from your clones? What is your template (e.g. scrape of colony in water, or mini-prep), and how much are you using for your PCR?
In Topic: cloning: band at different site than desired after RE digestion
06 January 2013 - 01:49 AM
Pangea, on 05 January 2013 - 01:18 AM, said:
It might be that your digestion of your Insert is not working out you have blunt ends. So it will just ligated several inserts. But i am not sure. And i hope you are not using BL21 for plasmid replication. Not recA mutation. But BLR have it or other K 12 strains. And are sure that you are not cutting into your plasmid?
