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Screening of chimeras for germ line transmission (GLT)

Article ID: 106
Last updated: 15 Dec, 2011
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The LacZ short range PCR detects the presence of the LacZ reporter in the cassette in the tm1a form of the EUCOMM / KOMP allele. This can be used as a rapid screen on F1 chimera progeny samples to detect likely germ line transmission. The WTSI genotyping assay is shown below. Once putative GLT has been detected, additional gene-specific assays and quality control tests can be performed (see www.knockoutmouse.org/kb/25/. for more information)

Assay

5' Forward Name
5' Forward Sequence
3' Reverse Name
3' Reverse Sequence
Product size
LacZ
LacZ_2_small_F
ATCACGACGCGCTGTATC
LacZ_2_small_R
ACATCGGGCAAATAATATCG
108bp

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