[Xenopus] pCS-GAP43-GFP sequence

Takuya Nakayama tn8t at virginia.edu
Wed Jun 11 15:13:03 EDT 2014


That's what I was about to point out too.
In case you wonder, here is the "T7" primer which we've been using for  
pCS2+ sequencing: 5'-AATACGACTCACTATAG-3' (just shorter version of  
original T7).

Takuya


On Jun 11, 2014, at 2:49 PM, Peter Klein wrote:

> Hi Jill,
>
> I agree with Sally. But the “T7" sequence in pCS2 is not the  
> standard T7 primer sequence, so sequencing facilities may not be  
> able to sequence from that end with their stock primers. I think you  
> need to provide the sequencing facility with a pCS2 appropriate “T7"  
> primer or use T3 (a little farther from the 3’ end). SP6 and T3 are  
> fine.
>
> I think this plasmid might be from Eddy DeRobertis lab, so you can  
> also check with them about it.
>
> Peter
>
>
> On Jun 11, 2014, at 2:02 PM, Sally Moody <samoody at gwu.edu> wrote:
>
>> Hi Jill,
>> We don't have that sequence, but in our experience it is better to  
>> send it out for sequencing rather than rely on somebody's map  
>> because things are not always what they seem. Since it is in pCS,  
>> you can use M13 primers or better yet SP6 and T7 primers for that  
>> plasmid.
>> Good luck, Sally
>>
>>
>> On Wed, Jun 11, 2014 at 1:46 PM, Jill Johnston  
>> <johnstoj at ucalgary.ca> wrote:
>> Hello,
>>
>> We have a construct called pCS-GAP43-GFP here and have no information
>> about it.  Does anyone have the sequence that could be sent to us?
>>
>> Thanks
>>
>>
>> Jillian Johnston
>> Research Associate
>> Dr. Sarah McFarlane lab
>> University of Calgary
>> Faculty of Medicine
>> Dept of Cell Biology and Anatomy
>>
>>
>>
>>
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>>
>>
>>
>> -- 
>> Sally A. Moody, Ph.D.
>> Professor of Anatomy and Regenerative Biology
>> George Washington University
>> School of Medicine and Health Sciences
>> 2300 I Street, NW, Washington, DC, 20037, USA
>> 202-994-2878
>> Editor-in-Chief:  genesis, The Journal of Genetics and Development
>> Editor: Principles of Developmental Genetics, 2nd edition
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>
>
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