1512 pT7A3-Phi29DNAPol

Cloning of 1512 pT7A3-Phi29DNAPol

Purpose: Cloning of 1512 pT7A3-Phi29DNAPol

Date: 2013-2-4---2013-2-

Notebook:0

Page:

Clone ID:1512

Experimenters: Shanyi

Results

subcloning

subclone Phi29DNAPol from 549 pET28b-Phi29DNAPol to 1279 pT7A3 using Nco1+ Xho1

	band 1	band 3
	1279pT7A3	549 pET28b-Phi29DNAPol
enzymes	Nco1+ Xho1

Colony Screening

Date: 2013-2-

Make miniprep and digest with Nco1+ Xho1

Result intepretation/Summary

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Conclusion

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Protein Expression

2013/3/11

Lane	Name	Protein Name	Clone ID	Host	Induction	Purification	pI/Mw
M	Invitrogen SeeBlue plus2
1	pT7A3-T4_RNA_Ligase_1	T4_RNA_Ligase_1	1049	5 ml, Nico21(DE3)plysN	0.2 mM IPTG  RT o/n	Ni beads	4.63 / 47485.21
2	pT7A3-MBP-(3C)-T4-RNA-Lig2	T4-RNA-Lig2	1123	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.11 / 39375.95
3	pT7A3-Klenow	Klenow	1501	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.11 / 71294.91
4	pT7A3-T4_RNA_Ligase_ 2_Truncated	T4_RNA_Ligase_ 2_Truncated	1503	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.14 / 29824.85
5	pT7A3-DNA-Pol-1	DNA-Pol-1	1508	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.40 / 103118.12
6	pT7A3-Phi29DNAPol	Phi29DNAPol	1512	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	8.61 / 66714.05
7	pT7A3-polyAPol	polyAPol	1513	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	9.67 / 53870.91
8	pT7A3-Klenow(3'->5'_exo–)	Klenow(3'->5'_exo–)	1537	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.77 / 67969.77
9	pT7A3-MBP-(3C)-Lambda-Exo	Lambda-Exo	1557	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.36 / 25925.62
10	pT7A3-MBP-(3c)-Endonuclease VIII	Endonuclease VIII	1572	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	7.77 / 29845.13