hMTMR11 (cDNA Clone SMINT2010144, FLJ39681, AK097000):

- Grew up plasmid (from NITE)

- PCR hMTMR4 sequence including Bgl II and Xho1 sites.

Primers:
Forward: 5’- t aAG ATC TCC ATG CCT CCC AGG GTC ACC TTC -3’ name=hMTMR11for.

length: 31
Tm: 66ºC
%GC: 55

Reverse: 5’- gc CTC GAG tta aca tct ggt tat cca gaa ggg a -3’ name=hMTMR11rev.

length: 33
Tm: 64ºC
%GC: 48

- After PCR, run product and recover 2.3 Kb band. Ligate into pCR4-Blunt-Topo (pCR-hMTMR11)

- Send for sequencing

- Subcloned into:
Prey pACT2…………………...with BamH1 and Xho1 (pACT2-hMTMR11)
Bait pFBT9………………….…with BamH1 and Sal1 (pFBT9-hMTMR11)

- For Co-IP, Co-IF:
Digest pCR-Topo-hMTMR11 with Bgl II and Xho1
pCMV-HA with Bgl II and Xho1
pEGFP-C2 with Bgl II and Sal1