1292 pET30b-PTD-DRBD-sso7d
Cloning of 1292 pET30b-PTD-DRBD-sso7d
[edit]Purpose: Cloning of 1292 pET30b-PTD-DRBD-sso7d
Date: August 1, 2012
Notebook:
Page:
Clone ID: 1292
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: August 1, 2012
Synthesized template via gene synthesize, expected size ~400 bp
Cut out band in lane 2, purify
Ligate , transform
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=== <u></u>Colony Screening<br> ===
Digested insert (see lane 3) and 368 vector (lane 4) with RE Kpn1 and Nde1, vector has too band bands, digest again, save insert for ligation over night
Date: August 2, 2012
Digested 368 vector (lane 3) with RE Kpn1 and Nde1, ligate together with digested insert from yesterday
Ligate together over night, transform tomorrow morning
Date: August 13, 2012
Screening came back mostly negative
Synthesized template via gene synthesize, expected size ~400 bp
Cut out band in lane 2, purify
Digested insert (see lane 3) and 368 vector (lane 4) with RE Kpn1 and Nde1, cut out and purify. LIgate together overnight
Colony Screening
[edit]Date: August 7, 2012
Digested MiniPrep with RE Kpn1 and Nde1
4 and 5 may be correct, maybe will digest again
Digested sample 4 (lane 1) and sample 5 (lane 2) again with RE Nde1 and Kpn1
Very faint band at expected size (~400)
Send sample 5 to sequencing
Date: August 20, 2012
Cannot tell, bad digestion
Date: OCO 26, 2012
Digested insert and vector with Nde1 + Xho1
Ligate , transform
Colony Screening
[edit]
sent to sequence miss one g
PCR
1117PCR-1118PCR 1422 1277 V15 1292 368 1139 1141 1147.JPG.JPG
[edit]
DIGEST pET30b-sso7d-KX 368 WITH Nde1 + Kpn1
SCREENING
Result intepretation/Summary
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Conclusion
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