Kamimura, S. et al. Mouse cloning using a drop of peripheral blood.
Mouse genomic clones used to generate the targeting construct were described earlier [20].
To confirm that the p53 pathway remained functional in FHL2−/− cell lines, we assessed the expression of p53 target genes at the whole-genome level by comparing gene expression profiles of three FHL2−/− cell lines with those of three immortalized wt MEF clones using Affymetrix Mouse Genome 430 2.0 microarray[5].
Additional filtering steps were required for de novo assembled transcripts to exclude false positives, so we mapped them against viral and ribosomal sequences and compared them with all known mouse, human, and rat ESTs and clones using BLAST.
Proof of principle for such "resurrection" was provided by an experiment in which mice were cloned using somatic cell nuclei derived from a mouse that was frozen for more than 15 years.
Human clones were preferentially sourced (ATAD2, BAZ2A, BRD4, CREBBP, SMARCA2 and ZMYND11), but where this was not possible, mouse clones were used (BRD3, BRD7 and TRIM24).
Where mouse clones were used, the amino acid sequences were >97% identical in the bromodomain region in all cases (see Additional file 2: Figure S2).
We performed sequence analysis of some of the cloned RT-PCR products to firmly establish the accumulation of rZH501-M847-A-type virus in rZH501-M847-G-infected mice; the numbers of the clones used for sequence analysis and the samples were: 20 clones from mouse 8b brain, 10 clones from mouse 6c spleen, 10 clones from mouse 7c brain, and 10 clones from mouse 7c spleen (see Fig. 8A).
The mouse RPTPρ cDNA was cloned using a combination of PCR and 5'-RACE.
As a positive control for the glycan-binding experiment, mouse PNGase, Ngly1, was also cloned using the primers listed in Table 3 to produce EcoRI sites at the both ends and with a 3' end FLAG-tag.
