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1677-pTCold-MBP-(3C) - Revision history
2026-10-18T16:44:02Z
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Paul.xu at 18:07, 10 April 2013
2013-04-10T18:07:54Z
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<p><b>New page</b></p><div>== '''Cloning of 1677-pTCold-MBP-(3C)''' ==<br />
<br />
'''Purpose:''' Cloning of 1677-pTCold-MBP-(3C) <br />
<br />
'''Date: '''2013-3-4---2013-3-7 <br />
<br />
'''Notebook:0''' <br />
<br />
'''Page:''' <br />
<br />
'''Clone ID:1677''' <br />
<br />
'''Experimenters:''' yingying <br />
<br />
== '''Results''' ==<br />
<br />
=== cloning ===<br />
<br />
[[Image:1677 pcr1 3-4-3013.JPG]] <br />
<br />
<br> <br />
<br />
pcr2&nbsp;:the same with 1611 pcr2 <br />
<br />
[[Image:1608+1610+1611+1612+1613 pcr 1-10-2013.JPG]] <br />
<br />
<br> <br />
<br />
pcr1+pcr2 <br />
<br />
[[Image:1673+1677 PCR 3-4-2013.JPG]] <br />
<br />
<br> <br />
<br />
[[Image:1677+1679+1680 clone 3-5-2013.JPG]] <br />
<br />
=== Colony Screening ===<br />
<br />
Date: 2013-3-7 <br />
<br />
Make miniprep and digest with BglII + Nde1 <br />
<br />
[[Image:1677 digest detect 3-7-2013.JPG]]<br> <br />
<br />
<br> <br />
<br />
== '''Result intepretation/Summary''' ==<br />
<br />
== '''Conclusion''' ==<br />
<br />
== '''protein screening''' ==<br />
<br />
[[Image:1567 1614 1673 1677 1692 1694 1697 1707 1711 protein screen 3-31-2013.JPG]] <br />
<br />
{| border="1" width="637" cellspacing="1" cellpadding="1"<br />
|-<br />
| lane<br> <br />
| name <br> <br />
| clone ID<br> <br />
| host<br> <br />
| induction<br> <br />
| colspan="2" | purification <br> <br />
| MW<br><br />
|-<br />
| 1<br> <br />
| pT7A3-T5-Exo<br> <br />
| 1567<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 21KD/5.95<br><br />
|-<br />
| 2<br> <br />
| pCold-TF-(3c)<br> <br />
| 1614<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| Ni beads<br> <br />
| 55.9KD/5.19<br />
|-<br />
| 3<br> <br />
| pET28b-Nde1<br> <br />
| 1673<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 44KD/8.41<br><br />
|-<br />
| 4<br> <br />
| pTCold-MBP-(3C)<br> <br />
| 1677<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 44KD<br><br />
|-<br />
| 5<br> <br />
| pCold-C(3C)-MBP-6H<br> <br />
| 1692<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 46KD/5.77<br><br />
|-<br />
| 6<br> <br />
| pT7A3-wtSM2<br> <br />
| 1694<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads, should use CBD beads<br> <br />
| 33KD/6.30<br><br />
|-<br />
| 7<br> <br />
| pCold-MBP-(3C)-TEV <br> <br />
| 1696<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 72KD/6.04<br><br />
|-<br />
| 8<br> <br />
| pCold-GST-(3c)-wt3C<br> <br />
| 1697<br> <br />
| 4 ml,NiCO21(DE3)pLysN<br> <br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 21KD/8.47<br><br />
|-<br />
| 9<br> <br />
| pT7A3-MBP-(E3C)-p53wt<br> <br />
| 1707<br> <br />
| 4 ml, <br />
BL21(DE3)Star, pRare2-pLysS <br> <br />
<br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 88KD/5.76<br><br />
|-<br />
| 10<br> <br />
| pET28b-hINFA2<br> <br />
| 1711<br> <br />
| 4 ml, <br />
BL21(DE3)Star, pRare2-pLysS <br> <br />
<br />
| 0.2 mM IPTG 25 oC o/n<br> <br />
| colspan="2" | Ni beads<br> <br />
| 21KD/5.95<br><br />
|}<br />
<br />
[[Category:Cloning]]</div>
Paul.xu