Methods Digest, Vol 19, Issue 3

Virash Gupta via methods%40net.bio.net (by virashkgupta At gmail.com)
Mon Dec 4 00:27:28 EST 2006


For Subject: sequencing prediction
20 is too much even to think of. even if you had gone for individual
ligation and sequenced the recombinant clones you were needed to sequence
only at the most 12x 3= 36 clones for to be scientifically acceptable. I
suggest you go for repeat ligation for rest of 8 clones and sequence
individual clones for each. Searching for sequence of these  8 clones stands
poor chance ( may be they are absent or present in very low number). In your
way, if lucky you may have next 8 clones sequenced the clones of your
choice, or if unlucky, you may not find even one in next 100 clones
sequence. In this science, if you face failure, it is always good to restart
from new end. This will help you in depression not taking over on you and
you can work with optimistic approach. By adopting this approach you can
also save time.
good luck.
Dr V K Gupta
Molecular Biology  lab
dept of Entomology
PAU Ludhiana-India.


On 12/3/06, methods-request At oat.bio.indiana.edu <
methods-request At oat.bio.indiana.edu> wrote:
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> Today's Topics:
>
>   1. Re: (no subject) (Mariana Coutinho)
>   2. ligation high (toyobo, japan) in the north america?
>      (dreadco At gmail.com)
>   3. sequencing prediction (Jinchun Wang)
>   4. Re: ligation high (toyobo, japan) in the north america? (Domo)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Sat, 2 Dec 2006 12:26:09 -0300 (ART)
> From: Mariana Coutinho <marianaloner At yahoo.com.br>
> Subject: Re: (no subject)
> To: methods At magpie.bio.indiana.edu
> Message-ID: <20061202152609.93373.qmail At web53904.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Hi
>
> You can raise polyclonal ascitys in mice which is easier to work with and
> results in and better antibody yeld.
>
> Article: development of transplatable ascitys tumours wich continuously
> produce polyclonal antibodies in pristane primed BALB/c mice immunized with
> bacterial antigens and complete Freund adjuvant.
> Cevenini, R.  Journal of Immunological Methods 140 (1991) 111-118
>
> regards
>
> mariana
>
> Dr Engelbert Buxbaum <engelbert_buxbaum At hotmail.com> escreveu:
> madhurima bhatnagar wrote:
>
> > Hello to all
> > I want to raise antibodies in mice for western blotting to detect
> > expression level in my transgenic plants.Can anybody suggest me any
> > references or protocols that can guide me in planning the experiments.
>
> Mice are too small to raise an antiserum, they are used for monoclonals.
> For serum, rabbits are better. Essential reading about immunological
> methods is
>
> @book{Har-88,
> AUTHOR= {E. Harlow and D. Lane},
> TITLE= {Antibodies --- A Laboratory Manual},
> YEAR= {1988},
> PUBLISHER= {Cold Spring Harbor Laboratory Press},
> ADDRESS= {Cold Spring Harbor},
> LANGUAGE= {engl}
> }
>
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> ------------------------------
>
> Message: 2
> Date: 2 Dec 2006 11:51:49 -0800
> From: dreadco At gmail.com
> Subject: ligation high (toyobo, japan) in the north america?
> To: methods At net.bio.net
> Message-ID: <1165089109.102278.21280 At f1g2000cwa.googlegroups.com>
> Content-Type: text/plain; charset="iso-2022-jp"
>
> hi,
>
> in japan, i used to use "ligation high" mix from toyobo for ligation.
> i wonder how i can purchase one in the north america?
> or do you know any equivalent products?
>
> thank you.
>
> lee
>
>
>
> ------------------------------
>
> Message: 3
> Date: Sat, 02 Dec 2006 22:29:17 -0600
> From: Jinchun Wang <jwang24 At mail.uh.edu>
> Subject: sequencing prediction
> To: "methods At magpie.bio.indiana.edu" <methods At magpie.bio.indiana.edu>
> Message-ID: <200612022229175463136 At mail.uh.edu>
> Content-Type: text/plain; charset=gb2312
>
> hi everyone,
>
> I have 20 different DNA duplexs in a mixture. After ligation,
> transformation, I got a plate of colonies. Now how can I predict how many
> colonies should be sequenced to find all 20 DNA?  If I already sequenced 100
> colonies and got 12 DNA, is there some model to calculate how many more
> should be sequenced to find others?
>
> Many thanks
>
> --------------
> Jinchun Wang
> 2006-12-02
>
>
>
> ------------------------------
>
> Message: 4
> Date: Sun, 3 Dec 2006 13:51:14 +0800
> From: "Domo" <rimask At yahoo.com>
> Subject: Re: ligation high (toyobo, japan) in the north america?
> To: methods At net.bio.net
> Message-ID: <457265d9 At news.starhub.net.sg>
>
> Is that good?
>
> I use MightyMix from Takara...
>
>
> <dreadco At gmail.com> wrote in message
> news:1165089109.102278.21280 At f1g2000cwa.googlegroups.com...
> > hi,
> >
> > in japan, i used to use "ligation high" mix from toyobo for ligation.
> > i wonder how i can purchase one in the north america?
> > or do you know any equivalent products?
> >
> > thank you.
> >
> > lee
> >
>
>
>
>
> ------------------------------
>
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> End of Methods Digest, Vol 19, Issue 3
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