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.

problem with faint bands

problem with faint bands - Protocols and Methods Forum

problem with faint bands - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

LinkBack Thread Tools Display Modes
Old 08-09-2006, 03:23 PM
bhairaja pandey
Posts: n/a
Default problem with faint bands


I have a problem with faint bands in AFLP. The bands are clear and strong around the higher marker base pairs (around 400 bp) however, the problem is that in the smaller size are not clear or very faint. I am doing Silver Staining for the developlopment of bands.


Get your email and more, right on the new Yahoo.com
Reply With Quote
Old 08-11-2006, 08:17 AM
Posts: n/a
Default problem with faint bands

I've never done Amplified Restriction Fragment Polymorphism but the
Invitrogen kit (11352010) manual or 10544013 for plants has some
trouble shooting tips:

[Only registered users see links. ]
[Only registered users see links. ] (for

Reply With Quote
Old 08-30-2006, 01:03 PM
Posts: n/a
Default problem with faint bands

I have quite a lot of experience with AFLP and I can say that the
thermocycler program is a critical parameter. At first I had similar
problems like you but then I optimized the cycler regime acording to a
manual by Applied Biosystems
[Only registered users see links. ].

It is very important to keep the ramp speed around 1deg/sec and to
extend the extension step to at least 1.5min (better 2min). The
touchdown protocol give better results e.g. first 10 cycles with 0.7deg
decrease each cycle.
Also, it is good to include an initial step 72deg for 2min in order Taq
to fill the adaptors and a final step 30min at 60deg. for the
transferase activity of Taq to add a terminal A or T.
I hope this helps


Reply With Quote

bands , faint , problem

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
Another contimation problem helpme Cell Biology and Cell Culture 3 05-18-2010 11:13 PM
How to avoid Nonspecific bands on multiplex PCR? Maddy PCR - Polymerase Chain Reaction Forum 3 02-24-2010 09:28 AM
blotchy, irregular bands rufus Western Blot Forum 7 09-29-2009 06:17 PM
Transfer problem Anais1979 Western Blot Forum 0 07-24-2009 03:55 AM
Simply put, MOVING DIMENSIONS THEORY is THE NEW MODEL: http://physicsmathforums.com drelliot@gmail.com Physics Forum 0 08-29-2006 06:32 AM

All times are GMT. The time now is 01:44 PM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2015, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.13335 seconds with 16 queries