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Gel |
Expected Results and Diagram |
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| Gene: 705 | Name: Capn5 | Family: Protease | Subfamily: Cysteine | Accession: Y10656 | GI: 2065241 |
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Gene: |
705 |
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ES
Line: |
1385 |
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Gene
Specific Oligo (GS): |
11621 |
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Gel |
Expected Results and Diagram |
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Gene: |
705 |
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ES
Line: |
1385 |
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Gene
Specific Oligo (GS): |
19587 |
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Gel |
Expected Results and Diagram |
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Legend : Genomic DNA from the
recombinant ES line was assayed for homologous recombination using polymerase
chain reactions (PCRs). Amplified DNA fragments were visualized by
ethidium bromide staining following agarose gel electrophoresis, as shown in
the gel image on the left. The test PCRs (a set of three lanes) employed the
gene-specific (GS) primer, which lies outside of and adjacent to the
targeting vector arm, paired in succession with one of three primers in the
insertion fragment. The "DNA sample control" employed a primer pair
intended to amplify a fragment from a non-targeted genomic locus. The
"positive control" employed the GS primer paired with a primer at
the other end of the arm. The table below the gel image lists the
primers (numbered) used in each PCR and the expected product sizes, in base
pairs (bp). The diagrams to the right depict the anticipated gel image
as well as the relative positions of the PCR primers. |
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Gene: |
705 |
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ES Line: |
1385 |
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Construct: |
4047 |
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Blot Number: |
2378 |
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Gene Side: |
5' |
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Legend : Genomic DNA isolated
from the indicated ES lines was digested with the indicated restriction
enzymes (determined to cut outside of the construct arms). The DNA was
analyzed by Southern hybridization, probing with a radiolabeled DNA fragment
that hybridizes outside of and adjacent to the construct arm. The
parent ES lines (negative controls) showed bands representing the endogenous
(wild-type) Gene #705 allele. In contrast, the ES#1385
line showed an additional band representing the targeted allele from the
expected homologous recombination event. "Additional"
denotes a secondary line that was derived from the same targeting vector but
was not used to generate mice. |
