Javascript Disabled Detected

You currently have javascript disabled. Several functions may not work. Please re-enable javascript to access full functionality.

phage434

Member Since 11 Sep 2004
Offline Last Active Yesterday, 05:38 PM

Find content

Community Stats

Group Global Moderators
Active Posts 1,805
Profile Views 24,663
Member Title Veteran
Age 65 years old
Birthday January 25, 1948
Gender
Male
Location
Cambridge, MA USA

About me

My research interests
mycoplasma genetics and metabolism

Contact Information

Website URL http://

130 Excellent

Friends

Latest Visitors

DeniseB
19 May 2013 - 05:11
blaszcz
03 May 2013 - 22:37
why_not_wheat
29 Apr 2013 - 13:30
yagmur
11 Apr 2013 - 01:00
Guest
29 Mar 2013 - 09:45

Posts I've Made

In Topic: Stain in electrophoresis is going from + to -

Yesterday, 03:44 PM

EtBr has the same problem. Either stain after running the gel, or add stain to the buffer at the positive electrode.

In Topic: "Easiest" sticky end combination

21 May 2013 - 06:27 AM

I like the Biobrick choice of EcoRI and PstI. Both are cheap, reliable, can be heat killed, work in the same buffer, are insensitive to E. coli methylation, and are available in high fidelity versions from NEB. They produce different overhangs: EcoRI a 5' overhang, and PstI a 3' overhang, so mis-ligation is unlikely. They also both cut near the end of dsDNA fragments with only a few bases outside the RE site.

In Topic: How to prepare serial dilutions of RNA or cDNA

16 May 2013 - 06:34 PM

Well, that is correct, it will damage the DNA. But in this application (and in almost all), you don't need very long strands -- they need only be long enough to contain both ends of your amplified region. Very long strands cannot be diluted accurately.

In Topic: How to prepare serial dilutions of RNA or cDNA

16 May 2013 - 02:41 PM

Genomic DNA is quite viscous and non-uniform in concentration within a sample, no matter how well you mix it. I'd suggest that with those samples you reduce the viscosity by mechanical shearing or cutting with rare cutters. Often forcefully expressing the sample through a small diameter needle several times will shear the DNA effectively.

In Topic: Ligation with small insert and large vector

16 May 2013 - 05:12 AM

5 ng of DNA should produce at least a million colonies if you have highly competent cells. Check your transformation efficiency with 10 pg of plasmid DNA, and calculate the efficiency. You should get 10**8 to 10**9 colonies per microgram of DNA.

Google Sign in options