PAAV-SMN1 302

Cloning of SMN1 to dsAAV-CB-MCS

Purpose: Gene synthesis of SMN1 and clone to dsAAV-CB-MCS .

Date: 2011-7-12

Notebook: Haiyan-1

Page:002995

Clone ID:302

Experimenters: Haiyan

Results

PCR amplify: primer SMN1-12; Pfu enzyme, 25 cycles;

Lane 1: Marker

Lane 2: 10 ul SMN1 pcr amplified product

Lane 3: dsAAV-CB-MCS, BglII+PstI

Lane 4: SMN1 PCR, BglII+PstI

Lane 5: SMN1 PCR, BglII+PstI

Ligate #3 with #4, #5

--Hylei 17:29, 13 July 2011 (EST)

Verification

Restriction Digestion

Lane 1: pAAV-Cre #1, digest with EcoR1 + Xho1

Lane 2: pAAV-Cre #2, digest with EcoR1 + Xho1

Lane 3: pAAV-Cre #3, digest with EcoR1 + Xho1

Lane 4: pDs-AAV-SMN1 #1, digest with BglII + Pst1

Lane 5: pDs-AAV-SMN1 #2, digest with BglII + Pst1

Lane 6: pDs-AAV-SMN1 #3, digest with BglII + Pst1

Lane 7: pAAV-Cre #1, digest with Nco1 + Xho1

Lane 8: pAAV-Cre #2, digest with Nco1 + Xho1

Lane 9: pAAV-Cre #3, digest with Nco1 + Xho, well leaks

Lane 10: pET30b , digest with Nde1 + Nco1

--Paul.xu 12:31, 17 July 2011 (EST)

Sequencing:

Result intepretation/Summary

•   pDs-AAV-SMN1 #1 and #2 may be good
  
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