Construction of pGADhnRNPA1-ΔM9;-ΔRG

˙Map

Insert: pEGhnRNPA1-ΔM9;-ΔRG (Cut BamHI,EcoRI and fill-in)

Vector: pGAD-GH (Cut BamHI and fill-in and CIP)

Check by cut NdeI+EcoRI or NdeI+HindIII

˙data

˙protocol

1.digestion----vector with BamHI and insert with EcoRI + BamHI

2.fill-in----using klenow (because the restriction sites of vector and insert are differient)

3.CIP----only vector to prevent it self-ligation

4.running gel, cut the bands & elute

5.ligation

6.transformation----the bacterial strain is JM101 (rAmp)

7.miniprep

8.check plasmid

(9.sequence)