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M.B.T's Content

There have been 10 items by M.B.T (Search limited from 17-October 18)


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#171760 solution

Posted by M.B.T on 19 November 2014 - 03:20 PM in General Lab Techniques

Perfect, thank u



#171758 solution

Posted by M.B.T on 19 November 2014 - 01:07 PM in General Lab Techniques

A thousand mM will give a M.

What about 250 mM, how many Mol/l is this?  Is this 0.25 M/L

 

What I try to ask was could I dilute a stock solution with certain molarity (e.g 1 M) to make another solution with another molarity (e.g 250 mM) or I can not dilute the molarity?

Thanks

  

 




#171752 solution

Posted by M.B.T on 19 November 2014 - 12:34 PM in General Lab Techniques

Hi everyone,

I need to ask something but I apology in advance if this kind of question was asked here before.

I need to make 10 ml of 5 X Loading buffer which required final concentration 250mM Tris HCL. I have 1 M Tis HCL stoke.

Using C 1 V1 = C2  V2 equation . I found out I need to take 2.5 ml of  1 M Tis HCL stoke to have 250 mM final concentration.

I am wondering what I did it right or I can not dilute molarity and I have to make a stock solution with 250 mM.

 

thank you

 

 

 




#166094 How to calculate MOI?

Posted by M.B.T on 17 March 2014 - 05:55 AM in General Lab Techniques

Hi every one,

 

I need to design an experiment and I need to Transduce my cells with vector dilution MOI 50.

Number of my cell is 5x10e5

Virus titter is 10e8

I am not sure how to calculate MOI, could you please help me with this one.

 

 

Cheers

 

 




#164695 maxi culture

Posted by M.B.T on 31 January 2014 - 11:27 AM in Molecular Biology

Hi everyone,
 
 




#163783 sequencing issue

Posted by M.B.T on 01 January 2014 - 05:08 PM in Bioinformatics and Biostatistics

ok, in this case should I re-streak from glycerol stock and sequence it again, am i right?

thank u for ur help




#163781 sequencing issue

Posted by M.B.T on 01 January 2014 - 04:44 PM in Bioinformatics and Biostatistics

How come mixed plasmid population, let me use a example to make it clear, let say i have sample No. 1, 

I aim to sequence two different parts of sample.

1: design two suitable primer for each part, so i have primer A and B

2: In one reaction, ~ 10 ul of sample 1 (DNA is product of mega prep) + primer A =  unredable sequencing result

3: in other reaction, ~ 10 ul of sample 1 + primer B= very nice result

 

any suggestion, still it could be mixed plasmid population. 




#163777 sequencing issue

Posted by M.B.T on 01 January 2014 - 07:33 AM in Bioinformatics and Biostatistics

Thank u for replay.

 

 

I was thinking of mix up the of cells of population, but the sample that I sent is product of mega pre and the thing is I used exactly same sample for two different sequencing reaction which one of them is absolutely OK and the other one is rubbish. I though if it is mixed up the cells of population then I should get bad result of both reaction because I used the same sample, am I right?

 

The same thing as well for G & C percentage, also is not a reign full of C&G.

 

If I want break it down, basically I got a same lentiviral vector backbone for 3 samples and the difference is only in their insert gene, so I have same promoter and other elements in all of them and to make sure about the both junction side during cloning I sent them for sequencing.

The sequencing of 5’ in all of them OK but the 3’ sequencing in 2 of sample is bad and the other one ok.

 

Any suggestion?




#163766 sequencing issue

Posted by M.B.T on 31 December 2013 - 07:43 AM in Bioinformatics and Biostatistics

Hi every one,

Happy New Year in advance.

 

 

I got three samples that they are same in promoter site and suppose to give exactly same sequence as well.

One of them has exactly the sequence that I expect but in the other two I can read the sequence only for 100 nucleotides, which isn’t enough to confirm the part that I am looking for. The strange thing is, the result become unreadable in exactly in same point in these two results.

What I thought might be some thing wrong by these two samples but on the hand I have sequencing of other part of these samples which is absolutely fine so the bad result couldn't be of poor sample.

The other possibility could be primer but the prime works and gave reading of 100 bp.

Any advise what could be the reason?

I thought something might be happened in company that I sent for sequencing.

 

 

 

 

Cheers




#163335 flag tag

Posted by M.B.T on 11 December 2013 - 02:52 PM in Biochemistry

hi every body.

 

 

I come cross a gene with two different format one is gene +flag tag on N-terminus and the other is gene + flag tag on C-terminus.

I am wondering to know what is reason to have flag tag on C or N-terminus, and what's benefit of this sites.

I know that N-terminus is at begging of protein and C-terminus before stop codon of protein. 

is any one can help with this, thank u.

 





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