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New Protocols for Isolating High- Molecular-Weight Genomic DNA

to determine the DNA concentration for each sample. The concentration was multiplied by the volume recovered to give the DNA yield.

cDNA (8 l) was packaged per the Transpack protocol, and an aliquot of the total DNA was plated on the Big Blue SCS-8 E. coli strain to determine the number of pfu per reaction.

dThe estimated total pfu recoverable was determined using the number of pfu per reaction multiplied by the number of estimated reactions achievable from the total volume of DNA recovered.

eThe pfu/ug DNA was determined by dividing the total recoverable pfu by the total yield of DNA.

fNR: none recovered.

g The average of all samples for mice, rats, and tissues combined (for each extraction technique). For comparison purposes, average values are for bone marrow and fibroblast only.

Blood samples were collected from Big Blue rats, while individual bone marrow samples were taken from both Big Blue mice and Big Blue rats. The Big Blue mouse embryonic fibroblast and Big Blue rat 2 embryonic fibroblast cell lines were also tested. Genomic DNA was isolated from two to five individual samples of each tissue type, using both dialysis and organic extraction methods. The results of the comparison are shown in Table 1. When using 5 to 7 ml of rat blood, the standard organic extraction protocol (Big Blue DNA isolation kit) failed to yield DNA, even after two attemp
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