Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Methods Digest, Vol 3, Issue 4

Methods Digest, Vol 3, Issue 4 - Protocols and Methods Forum

Methods Digest, Vol 3, Issue 4 - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 08-05-2005, 01:03 AM
TieQiao Wu
Guest
 
Posts: n/a
Default Methods Digest, Vol 3, Issue 4



Hi, everyone:

According to my experience to handle the Bio-Rad protean 2 system, I check the edges of glass plates carefully and discard those plates with seriously damaged edges. After using of glass plates I always use hot tape water to wash plates thoroughly. At the moment, we use another system called Hoefer Mighty small -multiple gel caster from Amersham Pharmacia. Using this system, you can cast upto 14 or 15 gels each time. Amersham also has multiple gel transferring system.

Cheers

Tieqiao

-----Original Message-----
From: [Only registered users see links. ]
[mailto:[Only registered users see links. ].indiana.edu]On Behalf Of
[Only registered users see links. ]
Sent: Friday, 5 August 2005 3:00 AM
To: [Only registered users see links. ]
Subject: Methods Digest, Vol 3, Issue 4


Send Methods mailing list submissions to
[Only registered users see links. ]

To subscribe or unsubscribe via the World Wide Web, visit
[Only registered users see links. ]
or, via email, send a message with subject or body 'help' to
[Only registered users see links. ]

You can reach the person managing the list at
[Only registered users see links. ]

When replying, please edit your Subject line so it is more specific
than "Re: Contents of Methods digest..."


Today's Topics:

1. suggestions for new PAGE apparatus (Dr. James J. Campanella)
2. RT-PCR (Johanna Fehling)
3. RE: suggestions for new PAGE apparatus (Jayakumar, R)


----------------------------------------------------------------------

Message: 1
Date: Wed, 03 Aug 2005 09:52:22 -0400
From: "Dr. James J. Campanella" <[Only registered users see links. ]>
Subject: suggestions for new PAGE apparatus
To: [Only registered users see links. ]
Message-ID: <6.1.0.6.0.20050803095118.03ab0db0@mail.montclair. edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

Hi everyone,

I am in the market to buy a new PAGE set-up. I refuse to buy Biorad again.
I have been burnt by their low quality equipment before. We have begun
pouring a large number of gels, and I am sick of wasting half my time time
trying to keep the casting system from leaking even as I'm pouring my gel.
There has to be something better out there than Biorad. I would like
recommendations on which company's apparatus will allow me to pour my PAGEs
every time without leakage.

Thanks for the help,

Jim

James J. Campanella,
Associate Professor
Department of Biology and Molecular Biology
Montclair State University
1 Normal Avenue
Montclair, NJ 07043

Alternate email address: [Only registered users see links. ]

Ph: 973-655-4097
Fax: 419-791-9834



------------------------------

Message: 2
Date: Wed, 3 Aug 2005 16:41:16 +0100
From: Johanna Fehling <[Only registered users see links. ].uk>
Subject: RT-PCR
To: [Only registered users see links. ]
Message-ID: <[Only registered users see links. ].uk>
Content-Type: text/plain; charset=US-ASCII; format=flowed

Does anyone know of an RT-PCR kit or protocol that would select for
long polyA tails? I am trying to do RT-PCR on protozoan cell cultures
contaminated with a mix of bacteria. Getting rid of the bacteria
doesn't seem to be an option (many things have been tried). Although
some bacterial transcripts are polyA'd, I think these tend to be much
shorter than in euk transcripts. I've been using the Strategene kit
"StrataScript Two-tube RT-PCR system with PfuUltra" but still seem to
be getting a lot of bacterial genes in my PCR products. I know polyU is
supposed to be more selective than polydT. Does anyone know of any kits
that use polyU, or any other tricks I might try?

Cheers,
Johanna

--------------------------------------------------------
Johanna Fehling
Postdoctoral Research Fellow
Department of Biology, Area 3
University of York
PO Box 373
York, YO10 5YW
United Kingdom

phone#: ++44 (0)1904 328633
fax#: ++44 (0)1904 328505
--------------------------------------------------------



------------------------------

Message: 3
Date: Wed, 3 Aug 2005 17:53:37 -0400
From: "Jayakumar, R" <[Only registered users see links. ]>
Subject: RE: suggestions for new PAGE apparatus
To: "Dr. James J. Campanella" <[Only registered users see links. ]>,
<[Only registered users see links. ].indiana.edu>
Message-ID:
<[Only registered users see links. ] swellpark.org>
Content-Type: text/plain; charset="us-ascii"


Hi.. I can see a lot of frustration directed at BioRad here.. :-)) I
too had problems earlier with Protean 2. After much trials and carefull
observation I found the problem and after that the plates have probably
leaked only 2-3 times in the past 3 years. The leak is most often due
to the following reasons. This is with my experience using mini protean
2 as well as the larger protein xi.
1. The bottom of the spacers don't exactly align with the plate edges
(run your finger through it check whether the spacer is flush with the
glass edges). Gel solutions can diffuse out through these spaces.
2. Glass plates often develop very small nicks and crack at the edges
due to use or due knocking against each other while washing. The gel
solution seeps out through these tiny nicks. Run a finger over the
edges to ensure that the edges are perfectly smooth.
3. Also check whether there is any solidified acrylamide sticking
between the spacers (to avoid this, clean and flush the casting
apparatus thoroughly under the tap IMMEDIATELY after use). Solidified
pieces of acrylamide can easily protrude between the spacers and the
glass causing the solution to leak.
4. These solidified pieces of acrylamide can also cause the glass
plates to crack when the screws are tightened. I have cut down on glass
breakage by more than 2/3rd by following a strict washing routine.
These will ensure a sure leak proof system and many more years of
happy casting experiences.
5. Also if the gel casting holders don't snap into place, change the
red foam pads (you can buy them separately from Biorad and stick them
into place). They get crushed with repeated use. Alternatively you can
try placing pads of whatman no3 paper between the red foam pad and the
grey cushion.

Best of luck
Jai

PS> I haven't found a better SDS-PAGE apparatus yet than bio-rad.

-----Original Message-----
From: [Only registered users see links. ]
[mailto:[Only registered users see links. ].indiana.edu] On Behalf Of Dr. James J.
Campanella
Sent: Wednesday, August 03, 2005 9:52 AM
To: [Only registered users see links. ]
Subject: suggestions for new PAGE apparatus
Importance: High

Hi everyone,

I am in the market to buy a new PAGE set-up. I refuse to buy Biorad
again.
I have been burnt by their low quality equipment before. We have begun
pouring a large number of gels, and I am sick of wasting half my time
time
trying to keep the casting system from leaking even as I'm pouring my
gel.
There has to be something better out there than Biorad. I would like
recommendations on which company's apparatus will allow me to pour my
PAGEs
every time without leakage.

Thanks for the help,

Jim

James J. Campanella,
Associate Professor
Department of Biology and Molecular Biology
Montclair State University
1 Normal Avenue
Montclair, NJ 07043

Alternate email address: [Only registered users see links. ]

Ph: 973-655-4097
Fax: 419-791-9834

_______________________________________________
Methods mailing list
[Only registered users see links. ]
[Only registered users see links. ]


This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you.



------------------------------

_______________________________________________
Methods mailing list
[Only registered users see links. ]
[Only registered users see links. ]

End of Methods Digest, Vol 3, Issue 4
*************************************


************************************************** ********************
This email and any files transmitted with it are confidential and
intended solely for the use of the individual or entity to whom they
are addressed. If you have received this email in error please promptly delete this email and notify
the system manager

************************************************** ********************


Reply With Quote
Reply

Tags
digest , issue , methods , vol


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Methods Digest, Vol 49, Issue 12 TieQiao Wu Protocols and Methods Forum 0 06-14-2009 11:28 PM
Methods Digest, Vol 49, Issue 12 TieQiao Wu Protocols and Methods Forum 0 06-14-2009 11:28 PM
Methods Digest, Vol 39, Issue 14 Large PCR p-GEM Jess Protocols and Methods Forum 0 08-23-2008 02:56 AM
Methods Digest, Vol 34, Issue 23 Protocols and Methods Forum 1 03-27-2008 10:57 AM
Methods Digest, Vol 17, Issue 23 Sayyari, Mohammad Protocols and Methods Forum 0 10-25-2006 09:11 AM


All times are GMT. The time now is 05:09 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.14777 seconds with 16 queries