PCR Primers for Transgenic Mice

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Line 12: Line 12:
*Transgene: tetO-Lac Z
*Transgene: tetO-Lac Z
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*5´ primer: GGC GTG TAC GGT GGG AGG
+
  *5´ primer: GGC GTG TAC GGT GGG AGG
-
*3´ primer: GTT GGG AAG GGC GAT CGG
+
  *3´ primer: GTT GGG AAG GGC GAT CGG
-
*PCR Fragment Size: 421 bp
+
  *PCR Fragment Size: 421 bp
   
   
 +
*Transgene: tetO-G-alpha 12
*Transgene: tetO-G-alpha 12
*5´ primer: CGA GAG CGG CAA GTC CAC
*5´ primer: CGA GAG CGG CAA GTC CAC

Revision as of 17:10, 29 June 2009

Conklin Lab Transgenic Mice

The Conklin Lab maintains several transgenic lines that utilize the tetracyclinetransactivator system. These lines will be made available on a collaborative basis and as our limited resources allow. (Jackson Lab should have manyof these lines soon).

Genotyping mice

Mice can be genotyped by PCR. After digesting a small piece of mousetail with proteinase K, a small alliquot of sample which contains genomic DNA is analyzed by PCR using transgene specific primers. Full protocol available.

All oligonucleotide sequences are shown as they should be ordered.

Tet-Responsive Lines (tetO)

  • Transgene: tetO-Lac Z
  *5´ primer: GGC GTG TAC GGT GGG AGG
  *3´ primer: GTT GGG AAG GGC GAT CGG
  *PCR Fragment Size: 421 bp

  • Transgene: tetO-G-alpha 12
  • 5´ primer: CGA GAG CGG CAA GTC CAC
  • 3´ primer: GCA GGC TCA GTC GGC ATG TAC
  • PCR Fragment Size: 245 bp

Transgene: tetO-G-alpha i2 5´ primer: GCA GAT CGA CTT TGC TGA CC

3´ primer: TCT GTC GGC ATG TAC TCC TC

PCR Fragment Size: 240 bp

Transgene: tetO-G-alpha q

5´ primer: TTG ACG TCC CCG ACT ACG CG 3´ primer: CTT TGG CCC CCT ACA TCG AC

PCR Fragment Size: 258 bp


Transgene: tetO-Ro1/Ro2 5´ primer (Go148): GTT GGG AAG GGC GAT CGG 3´ primer (Go147): GGC GTC TAC GGT GGG AGG PCR Fragment Size: 178 bp

Transactivator Lines Transgene: tTA 5´ primer: GCT GCT TAA TGA GGT CGG 3´ primer: CTC TGC ACC TTG GTG ATC PCR Fragment Size: 507 bp

Transgene: MH-tTA 5´ primer: TCA GAC CGA GAT TTC TTC ATC CC 3´ primer: CTC TGC ACC TTG GTG ATC PCR Fragment Size: ~1,000 bp

Transgene: MMTV-tTA 5´ primer: CTG ATC TGA GCT CTG AGT G 3´ primer: CTC TGC ACC TTG GTG ATC PCR Fragment Size: 595 bp

Protocols for genotyping the following mice can be found on the Protocol's Page:

TetO-Rs1 mice: Protocol to Genotype Rs1 Mice [1]


ColI(2.3)-tTA mice Protocol to Genotype ColI [2] (2.3)-tTA Mice: R. 20080528


NKX2-5-EmGFP (RP11-88L12 NKX2-5 EmGFP reporter mice) Protocol to Genotype NKX2-5 BAC Mice [3]

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