Search found 81 matches

by roniadam
Tue Sep 29, 2009 2:18 pm
Forum: Molecular Biology
Topic: Vector with no SV40poly A signal
Replies: 1
Views: 1198

Vector with no SV40poly A signal

Hi All:

I need to know what plasmid construct lacks the PolyA signal. I need this vector because I want to use an insert which contain the polyA signal so is there any vector I can purchase without the polyA signal.

Thank you
by roniadam
Tue Sep 08, 2009 3:03 am
Forum: Molecular Biology
Topic: Primer problem
Replies: 2
Views: 4854

Hi Jyaron:

There is no contamination in our reagents as this was excluded by using different primers which gave expected products except with using this pair. I reordered again the primers but the problem still is there :(
by roniadam
Sat Sep 05, 2009 3:10 am
Forum: Molecular Biology
Topic: Primer problem
Replies: 2
Views: 4854

Primer problem

Hi I designed a specific primers and blast it to amplify my gene. this primer amplifies a gDNA of the gene. I used dreamtaq kit and its protocol but unfortunately the expected product (2054 bp) was not there and what I got is several bands below and above my expected product size. Then I have used P...
by roniadam
Mon Jun 29, 2009 7:11 am
Forum: Molecular Biology
Topic: Lentivirus vector propagation
Replies: 4
Views: 3824

Lentivirus vector propagation

Hi

I bought a lentivirus vector containing alpha globin gene and I want to grow this vector. In what bug I should propagate it?
by roniadam
Wed Jun 17, 2009 1:01 pm
Forum: Human Biology
Topic: isolation of buffycoat
Replies: 0
Views: 1222

isolation of buffycoat

Hi everyone:

I want to isolate the buffycoat layer (MNCs) from a peripheral blood sample? How can I make to get a pure layer without red cell contamination. I have tried A Ficol solution but there was a contamination with the ficol particles so any one can help ???
by roniadam
Thu May 28, 2009 10:32 am
Forum: Molecular Biology
Topic: Big dye terminator
Replies: 1
Views: 2775

Big dye terminator

Hi :
I need to sequence a DNA. My primer conc. is 10 micromol and I need to convert it to 3.2 picomol to be able to set uo my sequencing reaction? how can I dilute my primer (10 micromol) to reach this small value of conc (3.2 picomol) ???? Anyone can explain to me in detail please?
by roniadam
Tue May 05, 2009 12:11 pm
Forum: Molecular Biology
Topic: cloning vector
Replies: 0
Views: 956

cloning vector

Hi All:

Can any one tell me if there is a vector that has a tag in which its poly A signal can be eliminated? I need this because I want to insert my own poly A signal. I was not able to find a vector. Any help please??
by roniadam
Mon May 04, 2009 10:14 am
Forum: Molecular Biology
Topic: Prep. of 50X TAE buffer
Replies: 8
Views: 33368

Thanx Canalon
I rally appreciate your kind help.
by roniadam
Sun May 03, 2009 6:06 am
Forum: Molecular Biology
Topic: Prep. of 50X TAE buffer
Replies: 8
Views: 33368

Thanks claudiupopa and Canalon

I am wondering why to add EDTA to 80 ml of MQwater rather than 100 ml? according to the protocol, I should add 100 ml of 0.5M EDTA. So how can I put it in 80 ml?
by roniadam
Sat May 02, 2009 9:29 am
Forum: Molecular Biology
Topic: Prep. of 50X TAE buffer
Replies: 8
Views: 33368

Prep. of 50X TAE buffer

Hi everyone: I am a new person to mol. bio. lab and I want to prepare 50 X TAE buffer. I did the follwoing: To make 1L of 50x TAE buffer: 242g Tris 100mL of 0.5M EDTA: This is prepared by (In 100 ml 1X PBS, Add 73.2 gram of EDTA) 57.1mL Glacial Acetic Acid Add enough MQ H2O to dissolve solids to fin...
by roniadam
Thu Apr 09, 2009 4:32 am
Forum: Molecular Biology
Topic: K562 cells for eryhtroid expression
Replies: 2
Views: 1707

Thanx LifeTein

Could you explain more as I am still confused.
by roniadam
Mon Apr 06, 2009 12:27 pm
Forum: Molecular Biology
Topic: K562 cells for eryhtroid expression
Replies: 2
Views: 1707

K562 cells for eryhtroid expression

Hi :

Can anyone explain to me why K562 cells (obtained from CML patient) are used for erythroid expression studies?
What is the relation between the CML and the erythroid cell lines?