MOLCULAR WEIGHT MARKERS

[Bruce Amsden]

pnorton at lac.jci.tju.edu (Pamela Norton) wrote:

>In article <1E7D931220 at oreo.uottawa.ca>, djohnson at OREO.CHEM.UOTTAWA.CA
>("Doug Johnson") wrote:

>> Could anyone suggest an alternative to HaeIII digested phiX174 as a
>> marker for Restriction enzyme or RAPD analysis? I would prefer to
>>  make my own, and in my hands a plasmid gives more DNA per cell
>>  than a virus. This is really an effort to save money. All suggestions
>are welcome.
>> 
>Doug,

>   We routinely use pBR322 digested with MspI for low MW markers, good for
>ca. 50-600 bp. We buy intact pBR and digest it ourselves, because the
>yields of plasmid are not great without chloramphenicol amplification of a
>large scale culture, but you certainly can grow it yourself. Oh, and the
>DNA labels up nicely with Klenow and dCTP, if you need hot markers. 

>   Feel free to mail if you need more info.

>      Pam Norton

>-- 
>Pamela A. Norton, Ph.D.          Assistant Professor of Medicine
>Thomas Jefferson University
>Philadelphia, PA 19107           p_norton at lac.jci.tju.edu


I use two digests of pUC18, which can be made by the bucketful.  Dde I
produces fragments of 910, 540, 426, 409, 234 and 166 bp.  Sau96 I
produces fragments of 1455, 616, 296, 222 and 75 bp.  The fragments
run well on agarose gels, although a couple of the bands run a bit
anomalously on polyacrylamide gels.

Bruce Amsden