Paul.xu at 23:25, 7 February 2019

2019-02-07T23:25:40Z

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Urgent Tasks

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| ID
| Clone Name
| Product Name
| Note
| Order ID
| Cat #
|-
| 162 
| pCold-MBP-(Tev)-GST-(3C)-Tat-NLS-iFLP-6H 
| Flp
| 
| 5148 
| EG-1002
|-
| 2108
| pCold-Tat-NLS-iFLP-6H 
| Flp
| 
| 5148 
| EG-1002 
|-
| 
| 
| 
| 
| 
| 
|-
| 2503 
| pCold-GST-VVTopo 
| Vaccinia Virus Topoisomerase I 
| 
|
5161

5154

| EG-167 
|-
| 
| 
| 
| 
| 
| 
|-
|
2291

|
pT7A3-SortaseAM-N6H

| Bifunctional Protein Ligase/Protease Sortase A 
| [http://www.excellgen.net/eln/index.php/212_429_2238_2197_2249_2287_2232_2291_2290_2294 expressed]
| 5159 
| EG-42 
|-
| 2232  
| pT7A3-SortaseAM-C6H 
| Bifunctional Protein Ligase/Protease Sortase A 
| [http://www.excellgen.net/eln/index.php/212_429_2238_2197_2249_2287_2232_2291_2290_2294 no expression]
| 5159 
| EG-42 
|-
| 
| 
| 
| 
| 
| 
|}

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| '''ID'''
| Status
| '''Name'''
| '''DNA Size'''
| '''Vector'''
| '''Method'''
| '''Progress''' 
| '''Date'''
| '''Enzyme''' 
| '''Expression'''
| '''Note'''
|-
| 3127 
| 
| pET28b-qBeta-qBeta-6XHis 
| 
| 
| 
| PCR qbeta and clone back into 3097
| 
| Nde1-HindIII
| C-His tag
| 
|-
| 812 
| 
| pET30b-phoA 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 938
| 
| PNGase F,
| 1145
|
<s>pET30b</s>

pT7A30

| Nde1-Xho1
| pJet1.2, sequencing is good, need to subclone into pET30b
| 
| 
| 
| C-His 
|-
| 939 pET30b-Tte-ExoIII
| 
| Tte-Exo III
| 780
|
<s>pET30b</s>

pT7A30

| Nde1-Xho1 
|
pJet1.2

sequencing is good, need to subclone into pET30b

| 
| 
| 
| C-His
|-
| 942 pET28b-pMBP-VVN
| 
| VVN endonuclease
| 716
| pET28b-pMBP 
| Nco1-Xho1
|
already in pJet1.2

need to check by digestion

| 
| 
| 
| C-His 
|-
| 3122
| 
| Crm197, secreted
| 1743
| pET30b 
| Nde1-Xho1 
| already in pJet1.2 
| 
| 
| 
| C-His
|-
| 931
| 
| PAK16D ligase
| 2267
| pET28b
| Nde1-HindIII-Xho1 
|
pJet1.2

fragment 2 is good

fragment 1 is also good

need to do digestions

seqeucning result: junk, E coli chromesome DNA

| 
| 
| 
| N-His
|-
| pT7A30-pMBP 945 
| 
| pT7A30-pMBP 
| 
| pT7A30
| Xba1-Nco1 
|
PCR from 1281 pMAL-p5E or 1282 pMAL-p5E-AvPAL

sequencing is good, completed

| 
| 
| 
| secreted to periplasm
|-
| pT7A30-6His-pMBP 947 
| 
| pT7A30-6His-pMBP 
| 
| pT7A30 
| Xba1-Nco1 
|
PCR from 1281 pMAL-p5E or 1282 pMAL-p5E-AvPAL (as template)

sequencing is good, completed

| 
| 
| 
| N-6Xhis tag MBP secreted to periplasm 
|-
| 944
| 
| pET30b-pfu-exo- 
| 
| 
| 
| 944 pET30b-pfu-exo digestion , sequencing result: not the right clone
| 
| 
| 
| 
|-
| 824 
| 0
| pET28b-VVTopo 
| 969 
| pET28b
| Nde1 + Xho1 
|
subclone from 832 pMiniT-VVTopo using Nde1 + Xho1

need to do PCR to confirm identity, if it is not the right clone, use 945 or 947 as vector

<s>824pCold-GST-VVTopoin pCold-GST vector, digestion , 824pT7A3-VVTpopin pT7A3 vector digestion</s>

in pET28 vector, sequenced

| 
| 
| 
| Customer's order #: 4988
|-
| 
| 
| 3107 pFastbac-Cel1 
| 
| 
| 
|
GGATCC ACCATGAAGTTCTTGGTGAACGTGGCGCTGGTGTTCATGGTCGTGTACATCTCCTACATTT ACGCGTGGGGCAAAGAAGGCCACTACGCCGTGTGTAAGATTGCGGAGGGCTTTTTGTCGGA AGACGCATTGGGAGCGGTCAAAGCCTTGTTGCCGGACTACGCGGAAGGCGACTTGGCAGCC GTATGTAGCTGGGCCGACGAGATCAGACACAACTTTCACTGGAGATGGTCGGGCCCACTGC ATTACGTCGACACGCCGGATTACAGATGCAACTACGAGTACTGCCGCGACTGTCACGACTT CAGAGGCCACAAAGACATTTGCGTCACGGGCGCGATATACAACTACACGAAACAATTGACG TCGGGCTACCACAACAGTGGCTCCGAGATTCGATACAACCTCACGGAGGCCTTGATGTTCC TCTCGCATTTCATTGGCGACGTGCACCAACCGCTGCATGTGGGCTTTACGGGCGATGAAGG CGGAAATACGATCATTGTCCGTTGGTACCGCAGAAAGACCAACCTCCACCACATATGGGAC AACATGATCATCGACTCGGCGTTGAAGACCTACTACAACAGCGACCTGGCCATAATGATCC AGGCGATTCAAAGAAACATCACCGGCGATTGGTCCTTTGACATCAGCAGCTGGAAGAACTG TGCCAGTGACGACACTGCTTGTCCGAACCTATACGCGTCGGAGAGCATCTCGTTGGCCTGT AAATTTGCCTACAGAAATGCCACCCCCGGTTCGACGCTGGGCGACGACTACTTCTTGTCGC GATTGCCGATTGTTGAAAAACGCCTCGCCCAAGGCGGTATTAGATTGGCCGCCACCTTGAA CCGTATTTTTGCCTCGCAACCGAAAATCTCGC GAAACACGAAGACAAGAGAG CGAGAAG ACGACGCCGGTAGACTACATCGAGTGGTCGCCAT TGCAACAGT TCAGCGGAAGCCACCACC ATCACCACCATCATCACGGC ctcgag CAT CAT CAT CAT CAT CAC TGATAA

AAGCTT

| 
| 
| 
| 
|-
| 995 
| 
| pCold-GST-CD133 
| 
| 3161 
| 
| no good
| 
| 
| 
| 
|-
| 994 
| 
| pMiniT2-LAMP-1_CD133 
| 
| 
| 
| no good
| 
| 
| 
| 
|-
| 993pCold-GST-M13-Gene8993 
| 
| pCold-GST-M13-Gene8 
| 
| 3161 
| 
| +993pCold-GST-M13-Gene8digestion
| 
| 
| 
| 
|-
| 992pCold-GST-V5992 
| 
| pCold-GST-V5 
| 
| 3161 
| 
| +992pCold-GST-V5digestion
| 
| 
| 
| 
|-
| 991pCold-GST-Flag991 
| 
| pCold-GST-Flag 
| 
| 3161 
| 
| +991pCold-GST-Flagdigestion
| 
| 
| 
| 
|-
| 990pCold-GST-HA990 
| 
| pCold-GST-HA 
| 
| 3161 
| 
| +990pCold-GST-HAdigestion
| 
| 
| 
| 
|-
| 989pCold-GST-Myc989 
| 
| pCold-GST-Myc 
| 
| 3161 
| 
| +989pCold-GST-Mycdigestion
| 
| 
| 
| 
|-
| 988 
| 
| 987 pCMVS-LAMP-1_CD133 
| 
| 192 
| 
| -
| 
| 
| 
| 
|-
| 985pCold-GST-3C-AP205-M13-g3p985 
| 
| pCold-GST-3C-AP205-M13-g3p 
| 
| 2047 
| 
| + 985pCold-GST-3C-AP205-M13-g3pdigestion
| 
| 
| 
| 
|-
| 984 
| 
| pT7A3-6H-SBP-SUMO-3C 
| 
| 1279 pT7A3 
| 
|  not good, sequencing is junk
| 
| 
| 
| 
|-
| 997
| 
| pT7A3-Qbeta 
| 
| 1279 pT7A3 
| 
| 997pT7A3-QbetaDigestion , sequenced , good, completed
| 
| 
| 
| 
|-
| 986
| 
| pT7A3-6His-NLS-Cas9-NLS-6xHis 
| 
| 1708 pT7A3-HTNCre 
| 
| 986pT7A3-6His-NLS-Cas9-NLS-6xHis digestion looks good, but need to do sequencing
| 
| 
| 
| 
|-
| 998 
| 
| pT7A3-6His-NLS-dCas9-NLS-6xHis 
| 
| 1708 pT7A3-HTNCre 
| 
| 
| 
| 
| 
| 
|-
| 999 
| 
| pT7A30-SssI 
| 
| 1916 
| 
| based on 1916 gene synthesis
| 
| 
| 
| 
|-
| 1000
| 
| pT7A3-6His-NLS-FnCpf1-NLS-6xHis 
| 
| 1708 pT7A3-HTNCre 
| 
| no good
| 
| 
| 
| 
|-
| 3210 
| 
| pT7A30-Sumo-Ov-ASP1-BCMA 
| 
| 1727 pT7A30 
| EcoR1+ Nde1
| 
| 
| 
| 
| 
|-
| 3218 
| 
| pT7A30-ULP1 
| 
| 1727 pT7A30 
| 
| PRC from 2209 ZM552-ULP1
| 
| 
| 
| 
|-
| 3191 
| 
| pT7A3-GST-Avitag 
| 
| 
| 
| sequencing: empty vector
| 
| 
| 
| 
|-
| 3192
| 
| pCMV5-PD-1-Avi-6H-BirA 
| 
| 
| 
| sequencing good, completed
| 
| 
| 
| 
|-
| 3166  
| 
| pCMV-mIgG2b-HC 
| 
| 
| 
| sequencing good, completed 
| 
| 
| 
| 
|-
| 3197
| 
| pT7A3-GST-BCMA 
| 
| 
| 
| no good 6/27/2018: sequencing: empty vector
| 
| 
| 
| 
|-
| 3194    
| 
| pCMV5-IL2-BCMA-(3c)-Fc-WPRE-OriP 
| 
| 
| 
| no good
| 
| 
| 
| 
|-
| 2503   
| 
| pCold-VVTopo 
| 
| 
| 
| subcloning  Nde1 -Xho1, digestion: not good, sequencing: junk 
| 
| 
| 
| 
|-
| 3218 
| 
| pT7A30-ULP1 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 2509
| 
| pT7A30-pMBP-Cid 
| 
| 947 pT7A30-6His-pMBP 
| 
| 
| 
| 
| 
| 
|-
| 3204 
| 
| pCMV5-OX40-Avi-6H-BirA 
| 
| 3192 pCMV5-PD-1-Avi-6H-BirA 
| 
| from 3203 pANT7_cGST-TNFRSF4 
| 
| 
| 
| 
|-
| 3199 
| 
| pCMV5-BCMA-Avi-6H-BirA 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 3196 
| 
| pCMV5-CD47-Avi-6H-BirA 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 3195 
| 
| pEF1-HTLV-LAMP-1-BCMA 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 3235 
| 
| pCMV5-IL2-MICA-(3c)-Fc-WPRE-OriP 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 3234   
| 
| pCMV5-IL2-PD-1-(3c)-Fc-WPRE-OriP 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 3233  
| 
| pCMV5-IL2-CD47-(3c)-Fc-WPRE-OriP 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 3232  
| 
| pCMV5-Avi-6H-BirA 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 2199   
| 
| pET30b-Ecoli-Muts 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 2204   
| 
| pET30b-Ecoli-Muts-D835R 
| 
| 
| 
| 
| 
| 
| 
| 
|-
| 129 
| 
| pL42-TthMuts 
| 
| 
| 
| 
| 
| 
| 
| 
|}
[[Category:To Do]] [[Category:Tasks]]

Cloning Tasks - Revision history

Paul.xu at 23:25, 7 February 2019