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zhenghj's Content

There have been 8 items by zhenghj (Search limited from 04-August 19)

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#4738 About fosmid librarty construction

Posted by zhenghj on 30 November 2003 - 05:35 PM in Molecular Biology

anyone here had used kit to construct fosmid library?I'v used a kit from EPICENTRE, but I can't get good result. So, is there any other companies that produced kit to construct library?

#4343 Subcloning transgene from a BAC clone

Posted by zhenghj on 29 June 2003 - 08:26 PM in Molecular Biology

To construct a subclone library from BAC. Perhaps u can construct a 15Kb library using phage vector.Is there any feature of ur gene for screening? If there's not,
u should disign a probe and do southen blot.Indeed I was trying to construct a 10Kb library using pUC18 as vector.Though it's not easy,but for BAC, it should be very easy since u need only a few clones.

#4315 A question about mRNA

Posted by zhenghj on 17 June 2003 - 08:14 PM in Immunology

I want to know ,when the content of a kind of antibody reaches the highest level
in the body, does it mean that the mRNA expressed it had reached the highest
level either?

#4216 sucrose gradient

Posted by zhenghj on 14 May 2003 - 12:51 AM in Molecular Biology

I have only done CsCl gradient centrifugation. I don't know the difference between
them.So you'd better see Molecular Cloning for help, I extract my sample in guidance of it and get good result.

#4211 Big Size of Plasmid Transformation

Posted by zhenghj on 09 May 2003 - 05:18 AM in Molecular Biology

where do u get the idea that electroporation needs large amount of DNA?
the efficiency of heat shock is much lower than electrooration, and for such a
big plasmid, elctroportion is a practical way. Don't care about the amount of
ur DNA.Good luck.

#4210 Stupid Ligation

Posted by zhenghj on 09 May 2003 - 02:42 AM in Molecular Biology

a 11 kb plasmid is not easy to transform, if u didn't get any clone, it may be the reason of low transformation efficiency.u'd better prepare a new electroporated competent cell.

#4209 Double strand cDNA sysnthesis

Posted by zhenghj on 09 May 2003 - 02:18 AM in Molecular Biology

Try Stratagene.It's difficult to check ur synthesis result in the process of synthesis.
Based on my experience,if ur RNA is good enough,go on ur synthesis until the step of ligation with vector,then u can check the amount of cDNA .

#4208 Restriction digest

Posted by zhenghj on 09 May 2003 - 02:09 AM in Molecular Biology

how about ur control? If u r digesting an enzyme site designed in PCR primer,
it's probably because that ur protection bases outside of the enzyme site is too

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