involve cutting a bulk DNA sample with restriction enzymes into a reproducible mixture of small fragments, size-separating these fragments by gel electrophoresis, transferring and binding the size-separated fragments to a solid support such as nylon, and then hybridizing a labeled DNA probe of analyte-complementary sequence across the support surface.
It has also reduced the number of Southern blots
required and decreased the turnaround time for patients with CGG repeat sizes up to 120 repeats.
Currently, laboratories either process every clinical sample on Southern blots
(because of the inadequacy of most FMR1 PCR tests) or reflex suspect samples to Southern blotting.
This is currently the most economical and versatile high throughput method of determining telomere length, although the values are not an actual kilobase pair length as compared to the data provided by Southern blots
. PBLs can be used as a valid measure of skin telomere length and this was reported by Friedrich in 2000 .
The Big Shot III hybridization oven is suited for all types of hybridization and incubation applications, including northern blots, southern blots
For Southern blots
, 10 to 15 [micro]g of genomic DNA was digested with HindIII restriction enzyme, electrophoresed in 0.8% (w/v) agarose gel, transferred onto Hybond-N+ (Amersham-Pharmacia Biotech) membranes, and fixed with a UV crosslinker (Stratalinker UV Crosslinker 1800, Stratagene, CA) as recommended in the manufacturers' instructions.
This was established by the failure of intron 1 and exon 9 primers to amplify from any of the BAC clones and by the lack of hybridization to the Southern blots
using a 2fc2 exon 9 probe.
For preparation of Southern blots
, total cellular DNA was isolated from overnight cultures as described previously (20).
The fcp probe also labeled Southern blots
of genomic DNA from Vaucheria litorea (not shown), not surprising since FCP's are always encoded by the algal nuclear genome (17).
The sequence of the p68 insert (Table 1) is 392 bp long, concordant with the size of the smallest bands on Southern blots
of genomic DNA hybridized with p68 (Fig.
The Big Shot III hybridization oven is suited to all types of hybridization and incubation applications, including northern blots, southern blots