Editing 938 pET30b-PNGase F

[[File:938 939 942 3122 Nde1 Nco1 Xho1.JPG|RTENOTITLE]]

[[File:938 939 942 3122 Xho1 HindIII.JPG|RTENOTITLE]]

12/27/2017

938 digestion with Nde1-Xho1, Fastdigest Buffer

[[File:938 939 942 3122 931 1855 digestion.JPG|RTENOTITLE]]

{| border="1" cellspacing="1" cellpadding="1" style="width: 681px;"
|-
| 1
| 2
| 3
| 4
| 5
| 6
| 7
| style="width: 54px;" | 8,9
| style="width: 55px;" | 10,11
| style="width: 71px;" | 13
|-
| 938&nbsp;PNGase_F
| <span style="color: rgb(0, 0, 0); font-family: Arial, Verdana, sans-serif; font-size: 12px; white-space: nowrap;">939 Tte-ExoIII</span><br/>
| 942 VVN<br/>
| 3122 CRM197<br/>
| 931, 1, PAK16D ligase&nbsp;<br/>
| 931, 2, PAK16D ligase<br/>
| 1855&nbsp; yPolyA-Pol<br/>
| style="width: 54px;" | 1540
| style="width: 55px;" | 2046
| style="width: 71px;" | 2046<br/>
|-
| Nde1-Xho1
| Nde1-Xho1<br/>
| <span style="color:#FF0000;">Nco1-</span>Xho1<br/>
| Nde1-Xho1<br/>
| Nde1-<span style="color:#FF0000;">HindIII</span><br/>
| <span style="color:#FF0000;">HindIII</span>-Xho1
| Nde1-Xho1
| style="width: 54px;" | Nde1-Xho1
| style="width: 55px;" | Nde1-Xho1
| style="width: 71px;" | <span style="color:#FF0000;">Nco1</span>-Xho1
|-
| pJet1.2
| pJet1.2<br/>
| pJet1.2<br/>
| pJet1.2<br/>
| pJet1.2<br/>
| pJet1.2<br/>
| pEC2-<br/>
| style="width: 54px;" | pET30b
| style="width: 55px;" | pET28b
| style="width: 71px;" | pET28b<br/>
|-
| 1145<br/>
| 780<br/>
| 716<br/>
| 1798<br/>
| 1174<br/>
| 986<br/>
| 1713
| style="width: 54px;" | there is no Xho1 site
| style="width: 55px;" | 1377
| style="width: 71px;" | ~1377
|-
| <br/>
| <br/>
| <br/>
| <br/>
| <br/>
| <br/>
| <br/>
| style="width: 54px;" | <br/>
| style="width: 55px;" | <br/>
| style="width: 71px;" | <br/>
|}

[[File:938 939 942 3122 931 1855 digestion Cutsmart.JPG|RTENOTITLE]]

==  ==

'''subclone of 938'''

Size:1145bp

V:1987 pT7A3

[[File:IMG 3672.JPG|RTENOTITLE]]

== Date:3/30/2018 ==

'''Miniprep digestion of 3122_938_939'''

3122:lane 1-4 lane 5&nbsp;:+ 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

[[File:IMG 0535.JPG|RTENOTITLE]]

'''redo Miniprep digestion of 3122_938_939'''

1:3122&nbsp; size:1743bp

2:3122pjet1.2 sequenced +

3:938&nbsp; size:1145 bp

4:938pjet1.2 sequenced +

5,6,7:939&nbsp; size:780 bp

8:939pjet1.2 sequenced +

Enzymes:NdeI+XhoI

[[File:IMG 0536.JPG|RTENOTITLE]]

'''938 miniprep test'''

1:938-1

2:938pJet1.2

Enzymes:EcoRV+XhoI

Size:552bp

3:938-1

4:938pJet1.2

Enzymes:NdeI+XhoI

Size:1071bp

[[File:IMG 0619.JPG|RTENOTITLE]]

== Date:4/5/2018 ==

'''938 subclone'''

'''Size:1145bp'''

'''Insert:938pJet1.2'''

'''Vector:1727 pT7A30'''

'''Enzymes:Nde1+Xho1'''

[[File:938subclone.JPG|RTENOTITLE]]

'''Miniprep digestion of 938_939_984'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| #
| Lane
| style="width: 34px;" | Enzymes
| style="width: 43px;" | Size
| Note
|-
| 938
| 1-4;5 is + control(sequenced 938pjet1.2)
| style="width: 34px;" | Nde1+Xho1
| style="width: 43px;" | 1145bp
| '''good'''
|-
| 939
| 6-9&nbsp;; 10 is + control(sequenced 939pJet1.2)
| style="width: 34px;" | Nde1+Xho1
| style="width: 43px;" | 780bp
| '''good'''
|-
| 984
| 11-14; 15 is + control(sequenced 982)
| style="width: 34px;" | Xba1+Nhe1
| style="width: 43px;" | 957bp
| need to redo
|}

[[File:Miniprepdigestion 938 939 984.JPG|RTENOTITLE]]

[[Category:Cloning]]