LaCava Research Wiki

Initiated September 2017

Hua Jiang's Journal

admin8th February 2022 at 2:20pm

February 3, 2022 - Making and testing new anti-ORF2 (9-7 and 5-5) beads

February 2, 2022 - Anti-ORF2 IP (MT646, MT647, LD564)

October 21, 2021 - MT646 S9.6 Tandem IP with RNase treatment before final elution

October 14, 2021 - MT646 and MT302 Tandem IP with anti-ORF1 and S9.6 beads

October 5, 2021 - Effect of RNase treatment on RNA/DNA hybrid and L1 RNP (MT646)

September 29, 2021 - anti-FLAG IP with MT646, MT647, LD564, MT302 and LD401

September 23, 2021 - Checking L1 expression of MT646, MT647 and LD564

September 8-15, 2021 - Conjugation of anti-ORF1 Dynabeads and attempt to reduce IgG leakage

September 2, 2021 - Concentrating and buffer exchange of anti-ORF1 (4H1)

August 30, 2021 - Testing Thrombin Sepharose Beads and Glutathione Resin

August 27, 2021 - Testing new MT302 powder

August 11-27, 2021- Transfection of HEK293TLD in suspension using PEI Max

August 18, 2021 - Testing new anti-ORF2 (clone 9-7)

August 16, 2021 - Making and testing anti-ORF1, anti-GFP and anti-FLAG beads

July 2021 - CRC 24-well screen_ 162 and 197

June 28, 2021 - Testing sonication conditions for 50mg powder

June 24, 2021 - CRC-197 anti-ORF1 IP with controls and beads titration

June 24, 2021 - Checking L1 expression of 9 L1 constructs

June 16, 2021 - pMT646 and pMT647 anti-FLAG IP

June 14, 2021 - Anti-ORF1 IP with senescent cells

June 9, 2021 - LINE-1 tandem prep with or without LaORF1 for EM

June 3, 2021 - Testing new anti-ORF2 beads and ORF2 dipeptides_2nd round

May 27, 2021 - Testing new anti-ORF2 beads and ORF2 dipeptides

May 27, 2021 - Testing new anti-ORF1 and mIgG beads

May 25, 2021 - Conjugation of anti-ORF1, anti-ORF2 and mIgG Dynabeads

May 24, 2021 - Anti-ORF1 IP with senescent cells

May 18, 2021 - SLE Neutrophils anti-ORF1 Western

May 14, 2021 - RRP6 purification followed by dialysis

May 13, 2021 - Effect of ZnCl2 and hUdT on CRC RNA quality

May 10, 2021 - Effects of ZnCl2 and hUdT on IP specificity and RNA quality

May 5, 2021 - Effects of ZnCl2 on IP specificity using CRC tissue

May 4, 2021 - Effects of ZnCl2 on IP specificity using different cell lines and tissue

May 3, 2021 - LD401 mIgG IP testing effects of ZnCl2 and new RNase inhibitor on IP specificity

April 30, 2021- N2102Ep anti-ORF1 IP with new RNase inhibitor and mIgG control

April 29, 2021- N2102Ep anti-ORF1 IP with or without new RNase inhibitor

April 27, 2021- RNA prep from CRC with new RNase inhibitor

April 19, 2021 - RNA prep with 3'-N-hydroxyurea-3'- deoxythymidine 5'-monophosphate

April 16, 2021 - Check SLE Western samples from Seattle

April 12, 2021 - Testing RRP6 powder

April 8, 2020 - Making and testing new anti-ORF1 beads

April 5, 2021 - Making and testing S9.6 and anti-ORF1 beads

April 1, 2021 - MT289 and N2102Ep Tandem IP using anti-ORF1, S9.6 Protein G and mIgG Epoxy control beads

March 30, 2021 - MT289, N2102Ep Tandem PO with S9.6 beads

March 24, 2021 - LINE-1 tandem prep_with or without LaORF1 for EM

March 22, 2021 - N2102Ep Tandem PO with S9.6 beads

March 9, 2021 - EXOSC10 (RRP6) gradient for EM

March 8, 2021 - L1 RNP preps to capture human auto-antibodies from serum/plasma_3rd test with 700ul serum

March 2, 2021 - N2102EP L1 screen in 24-well format (4x 6 conditions)

February 24, 2021 - Making and testing of mouse and rabbit IgG Dynabeads

February 17, 2021 - L1 RNP preps to capture human auto-antibodies from serum/plasma_2nd test

February 16, 2021 - Nucleic acids characterization in L1 RNPs_Collaboration with John Sedivy

February 10, 2021 - LD401 Tandem IP for EM_test ORF1 nanobody binding

January 28, 2021 - L1 RNP preps to capture human auto-antibodies from serum/plasma

January 7, 2021 - Large scale N2102Ep, NTERA anti-ORF1 and anti-ORF2 IPs for PacBio Sequencing

December 15, 2020 - N2102Ep, NTERA and CRC anti-ORF1 and anti-ORF2 IPs with controls

December 3, 2020 - N2102EP L1 screen in 24-well format

November 19, 2020 - Test ORF1 and ORF2 peptide for native elution after RNase A/T1 treatment

November 17, 2020 - N2102Ep anti-ORF1 IP with Nuclease treatments

November 10, 2020 - LD401 Tandem IP for EM with RNase A/T1 treatment on anti-FLAG beads

November 5, 2020 - RRP6 purification with BSA incubation

October 29, 2020 - Checking extraction conditions for 197N and 268N

October 28, 2020- RNA prep from CRC with DNase I treatment

October 22, 2020- RNA prep from CRC with and without EDTA

October 20, 2020 - N2102Ep and NTERA anti-ORF2 IP for Western

October 12, 2020 - N2102Ep anti-ORF1 and anti-ORF2 IPs

October 7, 2020- L1 endogenous cell line anti-ORF2 and rIgG IPs

October 5, 2020 - Summary of materials for CRC project

September 30, 2020 - LD401 Tandem for EM

September 29, 2020 - L1 endogenous cell line anti-ORF1 and mIgG for MS

September 23, 2020 - RRP6 purification with RNase A/T1 treatment for EM_092320

September 22, 2020 - Making and testing anti-ORF1 beads

September 17, 2020 - RRP6 purification with 300mM and 500mM buffer

September 16, 2020 - LD401 Tandem IP for EM

September 10-17, 2020 - RNA extraction and MS sample prep in endogenous cell lines following mock IP with mIgG

September 9, 2020 - RRP6 purification for EM

September 2-9, 2020 - Making and testing anti-FLAG beads with antibody from Marty

August 2020 - Removing 3x FLAG peptide from native elution

August 19, 2020 - RRP6 purification for EM

August 11, 2020 - Comparing anti-ORF1 monoclonal beads and LaORF1 nanobody beads_summary of experimental results

August 5, 2020 - anti-ORF1 IP with EDTA or MgCl2

July 30, 2020 - Optimizing AmAc concentration for protein precipitation

July 28, 2020 - Anti-ORF1 IP with N2102Ep and NTERA2

July 22, 2020 - CDI recombinant proteins

July 21, 2020 - AmAc/MeOH precipitation of protein from anti-ORF1 elution in organic phase

July 15, 2020 - Test Experiment of AmAc/MeOH precipitation_4th trial

July 7, 2020 - Testing anti-FLAG and uncoupled beads from ERIBA

July 6, 2020 - Anti-ORF1 IP with N2102Ep and NTERA2_070820

June 30, 2020 - Checking LaORF1 and LaG94 beads before and after DMP XL

June 29, 2020 - anti-ORF1 IP using anti-LaORF1 beads

June 29, 2020 - Test Experiment of AmAc/MeOH precipitation_3nd trial

June 22, 2020 - Test Experiment of AmAc/MeOH precipitation_2nd trial

June 18, 2020 - Test Experiment of AmAc/MeOH precipitation

June 17, 2020- CRC (159_185_174) anti-ORF1 IP

June 16, 2020 - Western Blots of Lars' samples

June 10, 2020 - CRC (162_185_197_197_268_284) anti-ORF1 IP

June 9, 2020 - Summary of anti-ORF1p RNA extractions by Lars

June 9, 2020 - Update on CRC project

Mar. 2, 2020 - SEC of ORF2 mAb (9-7)

Feb. 26, 2020 - Testing anti-ORF2 beads

Feb. 19, 2020 - SEC of ORF2 mAb (9-7)

Feb. 12, 2020 - Testing anti-ORF1_anti-ORF2 beads and RRP6-3xFLAG powder

Feb. 5, 2020 - Prepare whole cells and input samples for MS Using S-Trap Micro Spin Column Digestion Protocol

Jan. 29, 2020 - RRP6 purification

Jan. 22, 2020 - Capturing ORF2p with anti-ORF2 beads from N2102Ep

Jan. 7, 2020 - Capturing ORF2 with rabbit anti-ORF2 beads from N2102Ep and MT302

12/19/19 - PA-1, N2102Ep and NTERA2 anti-ORF1 IP

12/11/19 - Reuse anti-ORF1 for beads conjugation

12/04/19 - anti-FLAG, anti-ORF1 and anti-ORF2 beads titration test

11/12/19 - Making and testing LaORF2-12 and LaORF2-16 beads

11/06/19 - Making and testing LaORF2-12 beads

10/29/19 - Making and testing LaORF2 beads nanobodies

10/23/19 - Making and testing LaORF1 and LaG94 beads

10/22/19 - Making and testing anti-ORF1 and mIgG beads

10/17/19 - Antibody recovery from antibody mix after beads coupling

10/08/19 - anti-FLAG and anti-ORF2 beads titration test

10/07/19 - Conjugation of rabbit IgG Dynabeads

10/01/19 - Checking and recovery of anti-ORF2 (abcam, clone 9) antibody

09/30/19 - MT302 PO testing anti-ORF2 beads with or without XL

09/26/19 - Making_Testing anti-ORF2 beads

09/19/19 - Testing anti_ORF2 and LaORF2 beads

09/18/19 - Making anti-ORF2 (abcam Clone 9-7) beads with increasing AmSO4 concentration

09/16/19 - Making and testing abcam anti-ORF2 beads

09/14/19 - Testing abcam anti-ORF2 for Western

09/12/19 - abcam anti-ORF2 antibody info

09/09/19 - RNVC Testing – RNA Prep from MT302 and 162T

09/03/19 - 162T and 162N IP using PrG or PrA beads

08/26/19 - Making and testing anti-ORF1 beads

08/07/19 - Anti-ORF1 IP using PA-1 PC cells with crosslinked LaORF1 beads

08/05/19 - Orf2 (MT302) PO testing LaORF1 beads with or without XL

07/31/19 - Conjugation of LaORF1-5, LaG94-10 and anti-FLAG Beads

07/26/19 - BSA Standards in Urea and LDS

07/18/19 - Conjugation and testing anti-FLAG beads

07/10/19 - LD401_anti-FLAG and anti-ORF1 beads titration

07/08/19 - Conjugation of anti-ORF1 Dynabeads

07/02/19 - Prepare Whole Cell Samples Using S-Trap Micro Spin Column Digestion Protocol

06/28/19 - Sample Prep for PROTIFI

06/25/19 - Prepare IP Elution for MS Using S-Trap Micro Spin Column Digestion Protocol

06/17/19 - Testing new Abmart anti-ORF1 by Western

06/11/19 - LD567 Large Scale IP

06/07/19 - Making and testing anti-FLAG and anti-ORF1 beads

06/04/19 - Making and testing anti-FLAG and anti-ORF1 beads

05/28/19 - 162T and 162N anti-ORF1 IP_Sonication_Beads titration_Time course

05/16/19 - LD401, LD561 and pCEP-Puro anti-FLAG IP_400mg Scale

05/13/19 - LD401, LD561 and pCEP-Puro anti-FLAG IP

05/09/19 - L1 expression in LD401, MT302 and MT289

05/06/19 - ORF2/ORF1 Tandem to check LD401 powder

04/15/19 - MT302 and LD288 PO testing Llama anti-ORF2 antibody

04/08/19 - LD288, LD603 and pCEP-Puro anti-FLAG IP

04/03/19 - anti-ORF1 IP with MT302, PA1, HEK and HeLa cell lines

03/20/19- LD561 PO for MS using Gel Plugs or S-Trap

03/19/19 - Conjugating and testing anti-ORF1 beads

03/15/19 - L1 expression in MT289, MT302 and LD401 BB’s from Marty

03/13/19 - NCBP3 IP with GA Crosslinking

03/08/19 - CBP80 (NCBP1) anti-GFP IP_50mg scale

03/07/19- Conjugating Dynabeads with Llama anti-GFP antibody

03/06/19 - L1 expression in new MT302, LD401 and MT289 powder

02/27/19 - PA-1 and MT302 purification for MS

02/22/19 - Orf2 (MT302, PA-1) Test PO using anti-ORF2 Protein G beads

02/21/19 - Cross-linking of anti-ORF2 to Protein G beads

01/31/19 - Endogenous L1 Element Amplification (eLEAP) assay on MT302 and 162T / N

01/29/19 - eLEAP assay on PA-1 (50 and 100mg scale) with heat inactivation

01/22/19 - Endogenous L1 Element Amplification (eLEAP) assay on PA-1 and MT302 with heat inactivation

01/14/19 - Endogenous L1 Element Amplification (eLEAP) assay on PA-1 and MT302

01/09/19 - anti-ORF1 IP_Native Elution with ORF1 dipeptide

11/27/18 - Pullout with Ring 1B or mouse IgG beads using estrogen induced cells

11/19/18 - 162T and 164N anti-ORF1 PO for Western (Elution only)

11/16/18 - ORF2 (MT302) PO for EM_Urea

11/14/18 - 162T and 162N anti-ORF1 PO and Western of Lysate

11/12/18 - Isotope check for MT302H, MT289L and 302H/289LMix_SDS

11/07/18 - LD561 Purification for EM with Nucleases treatment

10/31/18 - ORF2 (MT302) test PO with different buffer conditions

10/30/18 - ORF2 (LD561) test PO with different buffer conditons

10/29/18 - Conjugating anti-Ring1B

10/24/18 - ORF2 (LD561)_PO for EM_with Nuclease treatment

10/18/18 - LD561 Test PO with Nucleases (Benzonase, RNase A/T1) treatment

10/17/18 - Isotope check MT302H, MT289L, 302H/289L mix

10/16/18 - ORF2 (LD561)_Test PO with Benzonase treatment

10/09/18- Anti-ORF1 IP with MT302 and PA-1 PC cells: titration of LaORF1 beads

10/03/18 - RRP6 purification for EM_Effect of detergent concentration of elution

09/27/18 - LINE1 glutaraldehyde crosslinking

09/24/18 - MT302 and PA-1 PC PO with anti-ORF1 beads

09/21/18 - anti-ORF1 beads survey

09/20/18 - Orf2 (MT302) PO with ORF1 beads titration

09/12/18 - Orf2 (MT302) PO with anti-ORF1 beads with or without XL

09/10/18 - Conjugating Epoxy beads with anti-ORF1 nanobodies and monoclonal antibody with and without XL

09/06/18 - RRP6 purification and concentration

09/04/18 - Orf2(MT302) Purification followed by RNAse treatment

08/29/18 - Orf2(MT302) Purification followed by RNAse treatment

08/23/18 - LD288 IP testing anti-FLAG beads

08/22/18 - RRP6 purification followed by gradient for EM

08/17/18 - RRP6 Purification with Native Elution

08/03/18 - Orf2 (MT302) Tandem IP using anti-FLAG and m6A beads with RNases treatment

08/02/18- Making anti-m6A Protein G beads_080218

07/31/18 - RRP6 Purification: Adhesive vs Suspension Cells

07/25/18 - LD288 anti-ORF1 PO (Epoxy beads without XL)

07/24/18 - anti-LaORF1 beads conjugation (Epoxy beads) with incresing amount of nanobody or AmSO4

07/18/18 - ORF2 (LD401) and RRP6 anti-FLAG PO

07/16/18 - Cross-linking (XL) of LaORF1 to Protein G or Protein A beads

07/13/18 - LD288 anti-ORF1 PO (Epoxy beads without XL)

07/12/18 - anti-LaORF1 beads conjugation (Epoxy beads)

07/11/18 - RNA Methylation in MT302 and LD401 Total RNA from Native Eluates

07/09/18 - RNA Methylation in MT302 and LD401 Native Eluates

06/29/18 - BSA Standard Stock for Gel

06/27/18 - RNA Methylation Quantification (MT302)

06/20/18 - Isotope check: LD401H and LD401H/LD259L

06/19/18 - Anti-ORF2 Protein G beads

06/18/18 - Orf2 (LD401, MT302) PO testing antibodies

06/13/18 - Effect of RNase treatment on RNA/DNA hybrid and Orf2 elution

06/12/18 - Effect of RNase H treatment on RNA/DNA hybrid

06/07/18 - Orf2 (MT302) Tandem IP using anti-FLAG and S9.6 beads with RNases treatment

06/05/18 - Orf2 (MT302) Tandem IP using anti-FLAG and S9.6 beads

05/31/18 - Orf2 (LD401) Tandem PO with S9.6 beads

05/30/18 - Anti-ORF2, S9.6, rIgG, mIgG Protein G beads

05/21/18 - Anti-ORF1 and anti-ORF2 Western in Human CRC tissues

05/15/18 - Testing S9.6 Protein G beads with Fluorescent Labeled Oligos and RNase treatment

05/14/18 - Testing S9.6 beads with fluroscent labeled oligos

05/09/18 - Testing S9.6 Antibody using LD401 Native Elution and Fluorscent Oligo

05/02/18 - Cross-linking of anti-DNA-RNA hybrid antibody (S9.6) to Protein G beads

05/01/18 - Test PO with anti-UPF1, NCBP1 and PABPC4 Protein G beads in HEK293 cells

04/30/18 - Cross-linking (XL) of anti-UPF1, NCBP1 and PABPC4 to Protein G beads

04/27/18 - Orf2 (MT302) Test PO using anti-ORF2 and anti-FLAG Protein G beads and anti-FLAG, anti-STAT3 Epoxy beads with or without XL

04/26/18 - Cross-linking (XL) of anti-ORF2 to Protein G beads

04/20/18 - Orf2 (LD401) Test PO using anti-ORF2 Protein G beads

04/19/18 - Cross-linking (XL) of anti-ORF2 to Protein G beads

02/12/18 - Testing S9.6 antibody against RNA/DNA hybrid

02/05/18 - MT302Heavy/MT289Light_I-DIRT 24-well Screen

02/01/18 - MT302Heavy/MT289Light_I-DIRT Manual Screen

01/24/18 - LD401Heavy/LD259Light mix_I-DIRT Screen

01/23/18 - Orf2 (MT302) PO with different amount of 3xFLAG peptide

01/19/18 - Orf2 (MT302) PO with different amounts of beads and peptide

01/17/18 - Orf2 (MT302) PO with different amount of anti-FLAG beads

01/12/18 - Orf2 expression in LD401 and MT302

01/12/18 - Orf2 (LD401) PO with post-PO Nuclease treatment

01/05/18 - Isotope check for L1 I-DIRT

01/03/18 - Protein Assay of L1 Heavy and Light Powder

12/07/17 - anti-KLF5 and anti-RELA PO and Western in HEK and HeLa

12/06/17 - Conjugating Dynabeads with CDI mouse mAbs

12/04/17 - anti-DACH1 PO and Western

11/16/17 - anti-FLAG and anti-NFKB1 Western

11/09/17 - NFKB1-3xFLAG test PO

11/07/17 - STAT3-3xFLAG Large Scale PO_150mM NaCl with control

10/31/17 - STAT3-3xFLAG Large Scale Purification_300mM NaCl

10/26/17 - LD401Heavy/LD259Light mix_I-DIRT 24-well Screen

10/16/17 - ORF2 (LD567 and LD624) anti-FLAG IP for RNA-seq

10/12/17 - ORF2/ORF1 Tandem for RNA-seq

10/05/17 - Total RNA prep with or without RNasin (LD401)

09/22/17 - ORF2 (LD567 and LD624) anti-FLAG IP for RNA-seq

09/20/17 - Making new anti-FLAG beads and test PO (LD401, 567, 624)

09/13/17 - ORF2/ORF1 Tandem for RNA-seq

Old data 04/02/14 - Orf2 (MT302) with 3x FLAG PO