JCA061606-Construction of key3 library 994

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PCR ca994F/ca996R on pSB1A2-J01129X (~2.2kb, XbaI/DpnI, Neb2)
Ligate, product is Lib994 library
Transform TG1/pSB3C6-J01122 cells


ca994F Forward XbaI EIPCR for key library
CCTATtctagaGACCCAAAAGCANGAGGNNNNNNTAGTNGGTGGTTAATGAAAATTAACTTACTTACTAG
ca996R Reverse XbaI EIPCR for lock and key
CGATGtctagaAGTGCTCAGTATCTCTATC


construction failed, potential new scheme:

PCR ca994F/G00101 on pSB1A2-J01129 (bp, XbaI/PstI, Neb2)
Sub into pSB1A2-R0040 (SpeI/PstI)
Product is Lib994

Alt new scheme: PCR ca994F/ca670R on pSB1A2-J01129 (bp, XbaI/AlwN1, Neb2)
Sub into pSB1A2-R0040 (SpeI/AlwNI)
Product is Lib994


construction of klenow extended 994Library
digest PCR product (XbaI/PstI, Neb2, small frag cleanup)
Paste into pJ23006-I13522 (XbaI/PstI, Neb2, Large-2334)
Product is pJ23006-I13522-994Library


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