Search found 6 matches

by Javedz2001
Sun Oct 11, 2015 5:19 pm
Forum: Molecular Biology
Topic: Restriction Digesion
Replies: 4
Views: 4856

Water bath is NOT recommended because after time the sample will evaporate and stick on the top of the cap. This will change the buffer composition. Always use PCR machine with heated lid or dry incubator.

Javed Siddiqi
by Javedz2001
Sun Oct 11, 2015 5:14 pm
Forum: Molecular Biology
Topic: TAE Buffer preparation
Replies: 1
Views: 4706

DO NOT add NaOH to any of the electrophoresis buffer. Strong acid or base can not be use to adjust the pH of electrophoresis buffer. It will have increase the current and heat up the runner buffer. Always use Acetic acid, boric acid, phosphoric acids for adjusting the pH. 50x TAE needs the following...
by Javedz2001
Sun Oct 11, 2015 5:01 pm
Forum: Molecular Biology
Topic: What an expression vector must have
Replies: 1
Views: 4055

The vector must have promoter (T7, lambda, lac etc. for E.coli), transcription start site and multiple cloning site for cloning your gene. Your target sequence must have ATG or should come from the vector. If you have to purify proteins then it should have some type of tag: his tag, etc. Good Luck: ...
by Javedz2001
Sun Oct 11, 2015 4:54 pm
Forum: Molecular Biology
Topic: How to perform gradient PCR?
Replies: 2
Views: 4508

The time will remain the same. Set up PCR as follows: FAST Cycling Conditions Temp (°C) Time (sec) Initial denaturation/Hot Start 95 30 Steps 1–3 are repeated through 40 cycles Step 1: 95 for 10 seconds Step 2: 54–70 (gradient) for 15 seconds Step 3: 72 15-60 seconds (depends on the length of your a...
by Javedz2001
Sun Oct 11, 2015 4:48 pm
Forum: Molecular Biology
Topic: My Multiplex PCR reaction works, but not my single rxn
Replies: 2
Views: 3737

Re: My Multiplex PCR reaction works, but not my single rxn

Multiplex PCR is very dependent on MgCl2 and dNTP concentration. Primer concentration also plays a big role. For individual PCR, try more primer and lower dNTP. You should also add 10% DMSO in the reaction. Lets us know if worked.

Good Luck: Dr Javed Siddiqi
by Javedz2001
Sun Oct 11, 2015 4:38 pm
Forum: Microbiology
Topic: NEW SPECIES - FECAL EATING BACTERIA
Replies: 3
Views: 6859

Re: NEW SPECIES - FECAL EATING BACTERIA

Hello: I saw your post about fecal eating bacteria. I am very interested in your finding. I have a biotech company and would like to collaborate with you on this project. Please reply here or send me your reply to [email protected]. Thanks.