I found a cost-effective way to create single-stranded DNA ladder in both low and high range (100-1000 bases, and 500-10000 bases). While making these neat ladders in hours, I am wondering what we can benefit from them. By Google, I found no ssDNA ladder is commercially available. However there are a few brands of ssRNA ladder which are used for RNA sizing and Northern blotting. We all know that RNA is much less stable compared to DNA. In addition, RNA is harder to be denatured. So, would people like to replace ssRNA ladder with the DNA one, if the latter could be easily made? The drawback I could see is that if one use the ssDNA ladder to size an RNA sample, the result would be off a bit (5%?) since RNA is heavier than DNA. If you guys have any suggestion on the application of the ssDNA ladder, please post it here. If anyone wants to try the ladder, please also let me know. Thanks!
0
6 replies to this topic
#2
mdfenko
-
- Active Members
-
- 2,240 posts
an elder
77
Excellent
Excellent
Posted 03 April 2012 - 11:59 AM
sizing primers
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#3
Trof
-
- Global Moderators
-
- 941 posts
Brain on a stick
66
Excellent
Excellent
Posted 03 April 2012 - 12:34 PM
100 bp primers would be pretty long.
Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.
I never trust anything that can't be doubted.
I never trust anything that can't be doubted.
#4
mdfenko
-
- Active Members
-
- 2,240 posts
an elder
77
Excellent
Excellent
Posted 04 April 2012 - 11:44 AM
true. maybe roller can make a ladder with even fewer bases (i have had a >60 base cassette synthesized and some microarrays use 60mers).
you can also size ssdna with which you may work.
you can also size ssdna with which you may work.
Edited by mdfenko, 04 April 2012 - 11:44 AM.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#5
hobglobin
-
- Active Members
-
- 5,249 posts
Growing old is mandatory, growing up is optional...
60
Excellent
Excellent
Posted 04 April 2012 - 12:07 PM
IMO when using a denaturing gel the standard dna ladder is also denatured and therefore ssDNA, or did I get something wrong?
Edited by hobglobin, 04 April 2012 - 12:49 PM.
One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.
#6
roller
-
- Members
-
- 3 posts
member
0
Neutral
Neutral
Posted 06 April 2012 - 10:11 AM
hobglobin, on 04 April 2012 - 12:07 PM, said:
IMO when using a denaturing gel the standard dna ladder is also denatured and therefore ssDNA, or did I get something wrong?
True when the size of DNA is small. For over 1k bp DNA ladder, it might be hard to completely separate the two strands even under denaturing conditions.
#7
roller
-
- Members
-
- 3 posts
member
0
Neutral
Neutral
Posted 06 April 2012 - 10:12 AM
mdfenko, on 04 April 2012 - 11:44 AM, said:
true. maybe roller can make a ladder with even fewer bases (i have had a >60 base cassette synthesized and some microarrays use 60mers).
you can also size ssdna with which you may work.
you can also size ssdna with which you may work.
Thanks for the suggestion.
