Search found 23 matches

by protold
Fri Dec 02, 2011 6:59 am
Forum: Microbiology
Topic: Viral budding
Replies: 1
Views: 3584

Re: Viral budding

Hola,early viral genes made that in the membrane of infected cells would appears protein molecules which facilitate anchoring of virions to new non infected cells spreading infections.when lysis of infected cell is produced , head of virions carries ,in their head ,membrane fragments with the viral ...
by protold
Thu Oct 20, 2011 6:12 am
Forum: Cell Biology
Topic: Sf9 Expression Kit that makes Blue Plaques with X-gal
Replies: 2
Views: 2125

Re: Sf9 Expression Kit that makes Blue Plaques with X-gal

Hola, BacNblue is the sole system wich gives blue recombinant plaques, but has the inconvenient that is incompatible with the rest whose have b-gal in the wild type virus so in a X-gal plating recombinant plaques will be white with a background less than 5% of non recombinant blue virus plates and h...
by protold
Mon Sep 05, 2011 5:49 am
Forum: Molecular Biology
Topic: Preparing buffer solns.
Replies: 6
Views: 5467

Re: Preparing buffer solns.

Hola in the case of acetic acid ,normality is the same that molarity so you can use 500x0.2=Vx17.46. Once you have the 10X TAE prepared , yes take a tent part and dilute with 9 parts of the final volume. This is a recipe of luckow and als manuals and I don´t remenber if you have to check the pH at a...
by protold
Mon Sep 05, 2011 5:39 am
Forum: Molecular Biology
Topic: Lipofectin/Labfectin lethality towards insect cells
Replies: 5
Views: 8278

Re: Lipofectin/Labfectin lethality towards insect cells

Hola, I used lipofectin and lipofectamin to transfec Sf9 cells and I haven´t big problems , I made minor volumes of transfection mix (100-200ul/p35mm)to save because if few viruses were formed giving more time of replication you could get enough population for amplification. But when I knew and use ...
by protold
Thu Aug 18, 2011 5:48 am
Forum: General Discussion
Topic: plasmid cut and uncut
Replies: 2
Views: 4174

Re: plasmid cut and uncut

Yes, plasmids show different mobility depending of their conformation. the fastest= supercoiled. above this linear digested and above these relaxed nicked circular form check any manual to confirm. Buena suerte
by protold
Thu Aug 11, 2011 6:07 am
Forum: Molecular Biology
Topic: Removing MBP tag
Replies: 12
Views: 14375

Re: Removing MBP tag

Hola, unless it seems silly here and I have seen some references about it in some forum, it´s better make digestion outside colum and after recharge the column with digestion and you will have your protein in the flowthrought, be carefull in pass the sample at low flow and be sure of don´t saturate ...
by protold
Wed Jul 20, 2011 5:39 am
Forum: Cell Biology
Topic: Neubauer haemocytometer-problem
Replies: 2
Views: 3550

Re: Neubauer haemocytometer-problem

hola,first looking the chamber without microscope you have to see the bars of divisions as an scratch on the two parts of chamber. after, under 40x objetive and playing with the condenser and power source to have less light you have to see the squares, but put any liquid between both glasses. Buena ...
by protold
Tue Jun 28, 2011 5:48 am
Forum: Microbiology
Topic: Identification of bacterial growth
Replies: 6
Views: 7548

Re: Identification of bacterial growth

Hola, Phenol red 15mg/l is used for growth indicator in culture media, when the pH drops bu sugar metabolism the colour changes to orange. But this is clear with solutions with carbon sources where the formed CO2 drops the pH. but if there isn´t sugars and the carbon is extracted from aminoacids by ...
by protold
Tue Jun 28, 2011 5:39 am
Forum: Cell Biology
Topic: Baculovirus amplification in sf9 cells.
Replies: 1
Views: 4430

Hola in my experience 4 days is the ideal time to colect virus passages to get titers of 10e8 to 10e9 pfu/ml. you have to suposse the first passages titers and see light infection symptoms when you harvest. for exemople you have to suposse a 10e4 for the titer of a transfection supernatant of 4 days...
by protold
Tue May 17, 2011 5:12 am
Forum: General Discussion
Topic: Biophysic
Replies: 1
Views: 1295

Hola check in VADLO.com. Buena suerte
by protold
Thu May 12, 2011 5:18 am
Forum: General Discussion
Topic: Molarities, concentrations and ratios
Replies: 3
Views: 2614

Hola first question : diluting 1/1000 a 50mM solution you obtain a 50uM sol.. So if you have 50ml of B and have to add A 1/1000 to have a 50uMsol of A, you have to add 1000 times less of volume so 50ul is the answer. 50ml/1000=50ul. Second one is less clair if sol C is 3 uM, is more diluted than 300...
by protold
Fri May 06, 2011 5:18 am
Forum: Molecular Biology
Topic: cloning
Replies: 10
Views: 8346

Hola, I´m sorry Jack Bean probably the exemple isn´t the best, I only wanted to say that there are silencious mutations. Excuse me, and have a good day