Comparison of bnl probes redux

--B. Burr and F.A. Burr

Last year Rick Johns and colleagues (MNL 64:56, 1990) reported variable results with 68 bnl probes. They reported that more than 25% gave too much background or were too faint to score. We were surprised by these results because one of the bases upon which we chose the probes was that they gave a strong signal. We went back to our original autoradiograms in which we had scored segregation in the TxCM or COxTx recombinant inbred families and found that all the probes in the Johns et al. worst category (D) gave respectable signals. Portions of these autoradiograms are reproduced here.

We published the specific conditions under which we used these probes in BurrB et al. Genetics 118:519-526, 1988. The major differences in our protocols appear to be that we used nitrocellulose membranes and nick translated our inserts.

Figure 1. Portions of original, working autoradiograms used to map loci detected by BNL clones that performed poorly for Johns et al. For each probe, five lanes of a segregating family are shown. For each panel the name of the locus is followed by the restriction enzyme used and the recombinant inbred family. (A)bnl3.04, BamHI, COxTx; (B)bnl4.24, BamHI, TxCM; (C)bnl5.02, EcoRI, TxCM; (D)bnl5.04, BamHI, TxCM; (E)bnl5.37, BamHI, COxTx; (F)bnl5.46, BamHI, COxTx; (G)bnl6.10, EcoRI, COxTx; (H)bnl7.49, BamHI, TxCM; (I)bnl8.04, BglII, COxTx; (J)bnl8.05, BglII, COxTx; (K)bnl8.10, EcoRI, COxTx; (L)bnl8.26, EcoRI, COxTx; (M)bnl8.33, BamHI, COxTx; (N)bnl8.39, BamHI, COxTx; (O)bnl9.08, BamHI, COxTx; (P)bnl10.12U, BamHI, TxCM; (Q)bnl10.13, EcoRI, COxTx; (R)bnl10.17, EcoRI, TxCM.

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