On Wed, Feb 1, 2012 at 6:42 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
> How many hours of cloning do you think you did? at 800bp * $0.30/bp ==
> $240... $240 / 16 hours = $15/hr... so at a low pay rate that's two
> days of work. Did you mess around more than 2 days before ligating the
> final repeat sequence into a vector?
>
Well, as I was inexperienced (been at least 2 years since I had done
cloning last) I think I made a few mistakes in the process. Otherwise
this would have been faster. When I had asked the question about how
to do this, it was just me that didn't know what was going on. The
boss already had the primers to do the deletion. I was trying to
figure it out myself.
And don't forget that we had the insert sequenced to double check that
it went in correct orientation and position. So another day and $22
How much DNA do the synthesis companies send back? We probably
extracted around 1 ug/uL (have to double check but it is high).
How do the companies synthesize? You send in the sequence what you
want, they put it thru their machine and send back some DNA?
I am guessing that our company was given this job because the customer
had some particular reason not to do this via commercial synthesis.
Cost was probably only one issue. I don't know much more.
> On Wed, Feb 1, 2012 at 6:24 PM, Jeswin <phillyj101@gmail.com> wrote:
>> Hey guys, the boss was able to get the insert as required and we just
>> did the maxi-prep and DNA purification today. Anyway, you all wondered
>> why it wasn't easier to get this synthesized instead of deleting
>> through PCR. I asked him and he said that the whole insert in which
>> the repeat occurs is 800bp long. This is too expensive to synthesize.
>>
>> Thanks for your help in this
>>
>> On Mon, Jan 23, 2012 at 8:51 PM, Jeswin <phillyj101@gmail.com> wrote:
>>> This sucks, I posted the primer in the above post. The repeat sequence is
>>>
>>> gggccaggtggtgcagggccaggtggtgcagggccaggtggtgcagggccaggtggtgcagggcccggtggtgcaggtccaggtggtgcaggtccaggtggtgcaggtccaggtggtgct
>>>
>>> I hate all this copy/paste. All the g,c,t,a make my eyes cross
>>> On Mon, Jan 23, 2012 at 8:45 PM, Jeswin <phillyj101@gmail.com> wrote:
>>>> On Mon, Jan 23, 2012 at 4:19 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
>>>>
>>>>> I meant what is the sequence that repeats... you gave two sequence
>>>>> snippets earlier, but they didn't seem to repeat 8 times in the .docx
>>>>> file you posted... not sure if you make some typos or I'm a victim of
>>>>> CTRL-F screwing up at line breaks (I was gonna throw the sequence into
>>>>> python/bioPython and do some crunching)
>>>>>
>>>>
>>>> My bad, I only posted the first line, forgot the rest:
>>>>
>>>> ggacgagctgtacaaggggccag
>>>> gtggtgcagggcccggtggtgca
>>>> ggtccaggtggtgcaggtccaggt
>>>> ggtgcaggtccaggtggtgctatg
>>>> gtgagcaagggcgaggagct
>>>>
>>>> The first few ends in "gca" but the rest end in "gct"
>>
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>
>
>
> --
> Nathan McCorkle
> Rochester Institute of Technology
> College of Science, Biotechnology/Bioinformatics
>
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