Editing
1513-pT7A3-polyAPol
Jump to navigation
Jump to search
Warning:
You are not logged in. Your IP address will be publicly visible if you make any edits. If you
log in
or
create an account
, your edits will be attributed to your username, along with other benefits.
Anti-spam check. Do
not
fill this in!
== '''Cloning of 1513-pT7A3-polyAPol''' == '''Purpose:''' Cloning of 11513-pT7A3-polyAPol '''Date: '''2013-2-25---2013-2- '''Notebook:0''' '''Page:''' '''Clone ID:1513''' '''Experimenters:''' Shanyi == '''Results''' == {| style="width: 281px; height: 66px" border="1" cellpadding="1" cellspacing="1" |- | <br/> | band 1<br/> | band 3<br/> |- | <br/> | 1279pT7A3<br/> | 1140 pTXB1-polyAPol<br/> |- | enzymes<br/> | colspan="2" | Nde1+ Xho1<br/> |} {| border="1" cellpadding="1" cellspacing="1" |- | <br/> |} === Colony Screening === Date: 2013-2- Make miniprep and digest with Nde1+ Xho1 [[File:1513digest.JPG|RTENOTITLE]] chose 1513-2 to do protein expression screening ,cannot express. [[File:1513-1DIGEST.JPG|RTENOTITLE]][[File:1513-1digest.JPG|RTENOTITLE]] == subcloning == subclone polyAPol from 1140 pTXB1-polyAPol to 1279 pT7A3 using Nco1+ Xho1 http://52.1.104.161/eln/images/4/4c/Digest_for_1513.JPG == Subcloning and Screening == Digested by Nde1 and Xho1 [[File:1944 1513 digestion.JPG]] == '''Result intepretation/Summary''' == *&lt;br&gt; == '''Conclusion''' == == Protein Expression == 2013/3/11<br> [[Image:1049 1123 1501 1503 1508 1512 1513 1537 1557 1572.jpg]] {| width="832" cellspacing="1" cellpadding="1" border="1" |- | Lane<br> | Name<br> | Protein Name<br> | Clone ID<br> | Host<br> | Induction<br> | Purification<br> | pI/Mw<br> |- | M<br> | Invitrogen SeeBlue plus2<br> | <br> | <br> | <br> | <br> | <br> | <br> |- | 1<br> | pT7A3-T4_RNA_Ligase_1<br> | T4_RNA_Ligase_1<br> | 1049<br> | 5 ml,<br> Nico21(DE3)plysN | 0.2 mM IPTG RT o/n<br> | Ni beads<br> | 4.63 / 47485.21 <br> |- | 2<br> | pT7A3-MBP-(3C)-T4-RNA-Lig2<br> | T4-RNA-Lig2<br> | 1123<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 5.11 / 39375.95 <br> |- | 3<br> | pT7A3-Klenow<br> | Klenow<br> | 1501<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads <br> | 5.11 / 71294.91 <br> |- | 4<br> | pT7A3-T4_RNA_Ligase_ 2_Truncated<br> | T4_RNA_Ligase_ 2_Truncated<br> | 1503<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 5.14 / 29824.85 <br> |- | 5<br> | pT7A3-DNA-Pol-1<br> | DNA-Pol-1<br> | 1508<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 5.40 / 103118.12<br> |- | 6<br> | pT7A3-Phi29DNAPol<br> | Phi29DNAPol<br> | 1512<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 8.61 / 66714.05 <br> |- | 7<br> | pT7A3-polyAPol<br> | polyAPol<br> | 1513<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 9.67 / 53870.91 <br> |- | 8<br> | pT7A3-Klenow(3'->5'_exo–)<br> | Klenow(3'->5'_exo–)<br> | 1537<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 5.77 / 67969.77 <br> |- | 9<br> | pT7A3-MBP-(3C)-Lambda-Exo<br> | Lambda-Exo<br> | 1557<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 5.36 / 25925.62 <br> |- | 10<br> | pT7A3-MBP-(3c)-Endonuclease VIII<br> | Endonuclease VIII<br> | 1572<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 7.77 / 29845.13 <br> |} <br> == Protein Expression 2 == 2013/3/29<br> SHANYI [[Image:1097 1138 1458 1494 1505 1513 1558 1562 1573 1593 1654NEW.jpg]]<br> {| width="832" cellspacing="1" cellpadding="1" border="1" |- | Lane<br> | Name<br> | Protein Name<br> | Clone ID<br> | Host<br> | Induction<br> | Purification<br> | pI/Mw<br> |- | M<br> | PageRuler Plus Prestained protein ladder<br> | <br> | <br> | <br> | <br> | <br> | <br> |- | 1<br> | pT7A3-T4-G32<br> | T4-G32<br> | 1097<br> | 5 ml,<br> Nico21(DE3)plysN | 0.2 mM IPTG RT o/n<br> | Ni beads<br> | 4.82 / 33506.43 |- | 2<br> | pT7A3-MBP-(3c)-MutM<br> | MBP-(3c)-MutM<br> | 1138<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 8.68 / 30290.07+43.6 <br> |- | 3<br> | pT7A3-KOD<br> | KOD<br> | 1458<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads <br> | 7.53 / 90022.42 |- | 4<br> | pT7A3-UDG<br> | UDG<br> | 1494<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 5.45 / 27174.73 |- | 5<br> | pT7A3-T7-Endo1<br> | T7-Endo1<br> | 1505<br> | 5 ml,<br> Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 9.54 / 17171.84 <br> |- | 6<br> | pT7A3-polyAPol<br> | polyAPol<br> | 1513<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 9.67 / 53870.91<br> |- | 7<br> | pT7A3-T7-Exo<br> | T7-Exo<br> | 1558<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 4.93 / 34502.20 |- | 8<br> | pT7A3-MBP-(3C)-Exo-1<br> | MBP-(3C)-Exo-1<br> | 1562<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 8.80 / 18143.72<br> |- | 9<br> | pT7A3-MBP-(3C)-RnaseH<br> | MBP-(3C)-RnaseH<br> | 1573<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n <br> | Ni beads<br> | 8.43 / <br> 17596.99 +43.6 |- | 10<br> | pT7A3-MBP-(3c)-E_coli_Endo IV<br> | MBP-(3c)-E_coli_Endo IV<br> | 1593<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2mM IPTG RT o/n <br> | Ni beads<br> | 5.43 / <br> 31479.53 +43.6 |- | 11<br> | pT7A3-T3 DNA ligase<br> | T3 DNA ligase<br> | 1654<br> | 5 ml, Nico21(DE3)plysN<br> | 0.2 mM IPTG RT o/n<br> | Ni beads<br> | 5.14 / 39350.70 <br> |} <br> [[Category:Cloning]]
Summary:
Please note that all contributions to ELN may be edited, altered, or removed by other contributors. If you do not want your writing to be edited mercilessly, then do not submit it here.
You are also promising us that you wrote this yourself, or copied it from a public domain or similar free resource (see
ELN:Copyrights
for details).
Do not submit copyrighted work without permission!
Cancel
Editing help
(opens in new window)
Navigation menu
Personal tools
Not logged in
Talk
Contributions
Create account
Log in
Namespaces
Page
Discussion
English
Views
Read
Edit
View history
More
Search
Navigation
Category:
Main page
To Do
Tasks
My ELN
Weekly Reports
Buffer Recipe
SOP
Protocols
Inventories
Projects
Products
Customers
Contacts
Suppliers
Vectors
Vector Map
Clones
Cloning
Sequencing
NGS
Protein Expression
Purchaseing
QC
293 Expression
CHO Expression
Sf9 Expression
Protein Purification
Stem Cells
Cytokines
Protein Kinases
GPCR
Cell Lines
Competent Cells
Viral Proteins
VLP
Baculovirus
AAV
Adenovirus
Lentivirus
Virus Titering
Antibodies
Transfection
ELISA
qPCR RT-PCR
ImmunoAssay
Multiplex BioAssa
Lateral_Flow
Conjugation
Compounds Synthesis
BioEngineering
GenomeEngineering
Services
Software
Ideas
Links
Knowledgebase
Employees
Company
Tools
What links here
Related changes
Page information
Navigation
Special pages