3154 pCMV5-HA-VH-mKappa

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2/17/2018

subcloning

1: insert, 

2: vector, top band

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miniprep digestion

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BamH1 digestion, to see if the clone has only one BamH1 site

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Redo

[edit]

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2/19/2018

Subclone HA heavy 3029-1 in pMiniT2 to 3130 pCMV5-HA-VL-mIgGkappa

to create an intermediate cloning vector which can be used for subcloning of Heavy chain 

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Miniprep digestion for 3154_3129

Digestion buffer:Fastdigest

Restriction enzymes:

3154:BamH I+ Kpn Iinsert length:527

3129:Kpn I+Not Iinset length:1034

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Result:seems good, chose 3129-1 to do sequencing

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