1569 pT7A3-MBB-(3C)-Exo-III

Cloning of 1569 pT7A3-MBB-(3C)-Exo-III

Purpose: Cloning of 1569 pT7A3-MBB-(3C)-Exo-III

Date: December 31, 2012

Notebook: 

Page:

Clone ID: 1569

Experimenters: Jennifer Truong

Results

Cloning

Date: December 31, 2012

Subclone from 1136 p8H-Exo-III (insert lane 3) using Nde1 + Xho1 and 1514 pT7A3-MBP-(3C) (vector lane 4)

Cut out fragments and ligate together and transform

Screening

Date: Jan 2, 2013

Digested MiniPrep with RE Xho1 and Nde1

See lanes 5-8

Only lane 5 (sample 1) has an insert, send to sequencing

Protein Expression

2013/3/20
SHANYI

Lane	Name	Protein Name	Clone ID	Host	Induction	Purification	pI/Mw
M	PageRuler Plus Prestained protein ladder
1	pT7A3-hTdT	hTdT	1509	5 ml, Nico21(DE3)plysN	0.2 mM IPTG  RT o/n	Ni beads	8.66 / 58536.26
2	pET28b-T7-DNA-Polymerase	T7-DNA-Polymerase	1533	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	6.04 / 79691.65
3	pET28b-T7-DNA-Pol-D28	T7-DNA-Pol-D28	1539	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.90 / 76333.85
4	pT7A3-MBP-(3C)-Exo-III	MBB-(3C)-Exo-III	1569	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.80 / 30969.17+43.6
5	pT7A3-T4RNA-Lig2-truncated-KQ	T4RNA-Lig2-truncated-KQ	1581	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.14 / 29796.84
6	pT7A3-MBP-(3c)-Ecoli-UDG	MBP-(3c)-Ecoli-UDG	1591	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	6.67 / 25693.23 +43.6
7	pT7A3-MBP-(3c)-newPhi29	MBP-(3c)-newPhi29	1617	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	8.61 / 66714.05 +43.6
8	pT7A3-MBP-(3C)-Exo-1	MBP-(3C)-Exo-1	1570	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.51 / 54484.77 +43.6
9	pT7A3-Klenow	Klenow	1501	5 ml, Nico21(DE3)plysN	0.1 mM IPTG RT o/n	Ni beads	5.08 / 71328.35
10	pT7A3-Klenow(3'->5'_exo–)	Klenow(3'->5'_exo–)	1537	5 ml, Nico21(DE3)plysN	0.1 mM IPTG RT o/n	Ni beads	5.77 / 67969.77
11	pT7A3-Ecoli_RecA	Ecoli_RecA	1529	5 ml, Nico21(DE3)plysN	0.2 mM IPTG RT o/n	Ni beads	5.09 / 37973.37