939 pET30b-Tte-ExoIII

12/27/2017

938 digestion with Nde1-Xho1, Fastdigest Buffer

1	2	3	4	5	6	7	8,9	10,11	13
938 PNGase_F	939 Tte-ExoIII	942 VVN	3122 CRM197	931, 1, PAK16D ligase	931, 2, PAK16D ligase	1855  yPolyA-Pol	1540	2046	2046
Nde1-Xho1	Nde1-Xho1	Nco1-Xho1	Nde1-Xho1	Nde1-HindIII	HindIII-Xho1	Nde1-Xho1	Nde1-Xho1	Nde1-Xho1	Nco1-Xho1
pJet1.2	pJet1.2	pJet1.2	pJet1.2	pJet1.2	pJet1.2	pEC2-	pET30b	pET28b	pET28b
1145	780	716	1798	1174	986	1713	there is no Xho1 site	1377	~1377

Date:3/27/2018

subclone of 939

Vector:1987 pT7A3

Date:3/30/2018

Miniprep digestion of 3122_938_939

3122:lane 1-4 lane 5 :+ size:1743 bp

938: lane 6-9 lane 10:+ size:1145 bp

939: lane 11-14 lane 15:+ size:780 bp

Positive control are insert-pjet1.2

The small band size of vector 1727:193 bp

redo Miniprep digestion of 3122_938_939

1:3122  size:1743bp

2:3122pjet1.2 sequenced +

3:938  size:1145 bp

4:938pjet1.2 sequenced +

5,6,7:939  size:780 bp

8:939pjet1.2 sequenced +

Enzymes:NdeI+XhoI

Date:4/4/2018

939 miniprep test digestion

1:939-1 1-4 enzymes:NdeI+AgeI

2:939-2 1-4 Size: 421bp

3:939-4

4:939pJet1.2

5:939-1 5-8 enzymes:XhoI+AgeI

6:939-2 5-8 Size:355bp

7:939-4

8:939pJet1.2 Results:all of them have bands

939 suclone

Size:780bp

Insert:939pJet1.2

Vector:1727 pT7A30

Enzymes:Nde1+Xho1

Miniprep digestion of 938_939_984

#	Lane	Enzymes	Size	Note
938	1-4;5 is + control(sequenced 938pjet1.2)	Nde1+Xho1	1145bp	good
939	6-9 ; 10 is + control(sequenced 939pJet1.2)	Nde1+Xho1	780bp	good
984	11-14; 15 is + control(sequenced 982)	Xba1+Nhe1	957bp	need to redo