Re: [DIYbio] large dna extraction

I did have what appears to be a 15kb band. I tried to purify that but
for some reason (age of kit, maybe?) I wasn't able to get dna out of
the gel. The project is on hold for now.

[1] Anyone know the shelf life of dna extraction beads (Qiaex II, not
sold anymore I think)?
[2] Also, I was told that the EtBr staining makes the DNA unsuitable
for sequencing. This is an invitrogen ion torrent machine. I have sent
pcr products for sequencing after gel purification without problems.
Is this machine different from the sequencers used by companies like
Operon?

On Tue, Feb 25, 2014 at 4:45 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
> I thought you said you had a 15kb band in the gel? Why don't you just cut it
> out and use that for sequencing?
>
> On Feb 25, 2014 9:04 AM, "Jeswin" <phillyj101@gmail.com> wrote:
>>
>> On Tue, Feb 25, 2014 at 9:42 AM, Koeng <koeng101@gmail.com> wrote:
>> > Do you need to get intact DNA? If not I usually just boil cells for a
>> > while to get DNA for pcr, which is pretty lazy but works quite well
>> >
>> It's for sequencing.
>>
>> Well, it turns out that the Qiagen Qiaex II kit (beads) to purify the
>> plasmid didn't give me any dna. Maybe the beads are too old. The gdna
>> that I collected off the gel (barely went into the gel) was purified
>> with the chargeswitch kit and gave a good amount of dna. Since we have
>> gdna, we can sequence the gdna and the total dna. Then, by
>> subtracting, we can determine the plasmid sequence. That's the plan,
>> anyway.
>>
>> --
>> -- You received this message because you are subscribed to the Google
>> Groups DIYbio group. To post to this group, send email to
>> diybio@googlegroups.com. To unsubscribe from this group, send email to
>> diybio+unsubscribe@googlegroups.com. For more options, visit this group at
>> https://groups.google.com/d/forum/diybio?hl=en
>> Learn more at www.diybio.org
>> ---
>> You received this message because you are subscribed to the Google Groups
>> "DIYbio" group.
>> To unsubscribe from this group and stop receiving emails from it, send an
>> email to diybio+unsubscribe@googlegroups.com.
>> To post to this group, send email to diybio@googlegroups.com.
>> Visit this group at http://groups.google.com/group/diybio.
>> To view this discussion on the web visit
>> https://groups.google.com/d/msgid/diybio/CAAhF0R%2BMRRsJyCT8i4D9GKbJwOsNpurRWhySYqmu4-gjRXFCpg%40mail.gmail.com.
>>
>> For more options, visit https://groups.google.com/groups/opt_out.
>
> --
> -- You received this message because you are subscribed to the Google Groups
> DIYbio group. To post to this group, send email to diybio@googlegroups.com.
> To unsubscribe from this group, send email to
> diybio+unsubscribe@googlegroups.com. For more options, visit this group at
> https://groups.google.com/d/forum/diybio?hl=en
> Learn more at www.diybio.org
> ---
> You received this message because you are subscribed to the Google Groups
> "DIYbio" group.
> To unsubscribe from this group and stop receiving emails from it, send an
> email to diybio+unsubscribe@googlegroups.com.
> To post to this group, send email to diybio@googlegroups.com.
> Visit this group at http://groups.google.com/group/diybio.
> To view this discussion on the web visit
> https://groups.google.com/d/msgid/diybio/CA%2B82U9KYB00_6h3EkBsuhj_t84COpj_ojQ6Uz1UMCRZmSrLORg%40mail.gmail.com.
>
> For more options, visit https://groups.google.com/groups/opt_out.

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at http://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/CAAhF0RJNsay3VVg3fULfor6QtXW3DHs5Vv%2B%3DFXGfAKQqR0SB3w%40mail.gmail.com.
For more options, visit https://groups.google.com/groups/opt_out.

  • Digg
  • Del.icio.us
  • StumbleUpon
  • Reddit
  • RSS

0 comments:

Post a Comment