Question: How to sequence a gene along with its 3' and 5' UTR
0
gravatar for Shahzad
2.6 years ago by
Shahzad10
Pakistan
Shahzad10 wrote:

Dear All I have a gene transferred into a plant (cotton) chromosome. I want to know the upstream and downstream DNA sequences present with this gene. Please guide me the possible procedure/s by which I can get this information and specially is there any possibility that NGS can help in this matter. If anymore information is needed please ask.

BEST REGARDS

utrs sequence gene • 696 views
ADD COMMENTlink modified 2.6 years ago by Carlo Yague4.4k • written 2.6 years ago by Shahzad10
2

Do you want the flanking DNA regions (not part of the transcript) or the UTRs which are part of the gene and are transcribed ?

ADD REPLYlink written 2.6 years ago by Carlo Yague4.4k

I want the sequences of flanking DNA regions(upstream and downstream) to know the place of this Gene in a chromosome.

ADD REPLYlink written 2.6 years ago by Shahzad10
2

If you know the sequence of your inserted gene, you could try "cycle sequencing" by designing a single primer at the end of your gene that will read into the flank (and then do the same for the start).

ADD REPLYlink written 2.6 years ago by christopher medway440

Thank you all for these wise suggestions.

ADD REPLYlink written 2.6 years ago by Shahzad10
1
gravatar for Carlo Yague
2.6 years ago by
Carlo Yague4.4k
Belgium
Carlo Yague4.4k wrote:

NGS can definitely help if you do genome sequencing. This thread may give you some ideas on how to proceed with the data.

ADD COMMENTlink written 2.6 years ago by Carlo Yague4.4k

please also let me know if there is Any other technique which can earmark the transformed gene's position in chromosome so I can get the up and downstream DNA sequences?

ADD REPLYlink written 2.6 years ago by Shahzad10
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