CBM UniTrap Project

UniTrap UNI7064

Trapped Gene	Mphosph6 (ENSMUSG00000031843)
Vector Insertion	Chr 8: 120318572 - 120322943
Public Clones	RRZ074 (baygenomics)
Private Clones	OST59329 (lexicon)
Severity of mutation (?)	Insertion after 34% of polypeptide chain

Suggested RT-PCR (?)

Control PCR for the trapped allele
Upstream Exon ENSMUSE00000320725 (Chr8:120322944..120323056 -)				Vector Sequence
Forward Primer	Location	Tm	%GC	LacZrt Reverse Primer	Location
CTGGACTCGGAAACCAAGAA	Chr8:120323022..120323041	60.22	50	TGGCGAAAGGGGGATGTG	Gene trap vector


Control PCR for the wild-type allele
Upstream Exon ENSMUSE00000320725 (Chr8:120322944..120323056 -)				Downstram Exon ENSMUSE00000320718 (Chr8:120318481..120318571 -)
Forward Primer	Location	Tm	%GC	Reverse Primer	Location	Tm	%GC
CTGGACTCGGAAACCAAGAA	Chr8:120323022..120323041	60.22	50	TCAGGATTAAACCCCCTGAA	Chr8:120318466..120318485	59.36	45

Other possible Exon-Primers for RT-PCR (?)

	Exon ID	Exon location	Forward Primer	Primer location	Tm	%GC
upstream	ENSMUSE00000320735	Chr8:120325752..120325829	TCGGAGCGCAAGACTAAGTT	Chr8:120325777..120325796	60.15	50
upstream	ENSMUSE00000320725	Chr8:120322944..120323056	CTGGACTCGGAAACCAAGAA	Chr8:120323022..120323041	60.22	50
* Putative Vector Insertion (Chr 8: 120318572 - 120322943) *
	Exon ID	Exon location	Reverse Primer	Primer location	Tm	%GC
downstream	ENSMUSE00000320718	Chr8:120318481..120318571	TCAGGATTAAACCCCCTGAA	Chr8:120318466..120318485	59.36	45
downstream	ENSMUSE00000213654	Chr8:120316579..120316676	TCGACCTCTACCGTCTCGTC	Chr8:120316584..120316603	60.41	60
downstream	ENSMUSE00000320701	Chr8:120315545..120316218	ACCAGTACCGACGACAGGAG	Chr8:120315988..120316007	60.17	60

Suggested Genomic-PCR (?)

	Forward	Forward location	Tm	%GC	Reverse name	Reverse	Reverse Location
PCR 1	CGAGCACTGGTACCTGGATT	Chr8:120322964..120322984	60.13	55	L232	GATGTGCTGCAAGGCGATTA	Gene trap vector
PCR 2	CGAGCACTGGTACCTGGATT	Chr8:120322964..120322984	60.13	55	LacZ	CCAGGGTTTTCCCAGTCACG	Gene trap vector

Come back to gene ENSMUSG00000031843