PMID:35293864 #8
Description
Submitter 1 full name
Henrique Filipe Oliveira Leonardo
Submitter 1 ORCID
No response
Submitter 1 GitHub username
hfoleonardo
Submitter 2 full name
No response
Submitter 2 ORCID
No response
Submitter 2 GitHub username
No response
Submitter 3 full name
No response
Submitter 3 ORCID
No response
Submitter 3 GitHub username
No response
PMID
PMID:35293864
Title
Microtubule rescue at midzone edges promotes overlap stability and prevents spindle collapse during anaphase B.
Cloning Detail Level
Cloning information present, but not sufficient to reproduce
Plasmids and Alleles
- Plasmid pSR176 (pFA6a-natMX6-Pnmt81) (Addgene #39280)
- natMX6:P81nmt1-cls1-3xGFP:kanMX6
- Plasmid pSR174 (pFA6a-natMX6-P1nmt1) (Addgene #39282)
- natMX6:Pnmt1-cls1-3xGFP:kanMX6
- pKS394 (pFa6a-kanMX6-P1nmt1-mCherry)
- kanMX6:P1nmt1-mCherry-ase1
- pKS395 (pFa6a-kanMX6-P41nmt1-mCherry)
- kanMX6:P41nmt1-mCherry-ase1
- pML1 (pFa6a-mCherry-ase1:kanMX6)
- mCherry-ase1:kanMX6
- Plasmid pFA6a-mEGFP-KanMX6 (Addgene #87023)
- nem1Δ:NatMx6
Cloning Techniques Used
- Gibson Assembly
- Golden Gate
- Gateway Cloning
- CRISPR/Cas9
- Homologous Recombination
- Traditional Restriction Enzyme Cloning
- PCR Cloning
- Overlap Extension
- In-Fusion Cloning
- De-novo DNA synthesis
- Other (please specify in notes)
Cloning Text (if available)
"### Production of S.pombe mutant strains
All used strains are isogenic to wild-type 972 and were obtained from genetic crosses, selected by random spore germination and replicated on plates with corresponding drugs or supplements. Gene deletion and tagging was performed as described previously (Bähleretal.,1998), following (Kawaietal.,2010). All strains, oligonucleotides and plasmids used, and how they were made are described in the Supplementary File 1."
elife-72630-supp1-v2.xlsx
elife-72630-v2.pdf
Cloning Strategies
cloning_strategy2.json
cloning_strategy.json
Missing information (if any)
The plasmids pKS394 (pFa6a-kanMX6-P1nmt1-mCherry) and pKS395 (pFa6a-kanMX6-P41nmt1-mCherry) are from Prof Ken Sawin's lab, thus their sequences were not available online.
The TP5771 strain mentioned in the Supplementary File 2 was not described or mentioned anywhere else.
Present information (if any)
The paper had all the strains and its identifications, although I didn't know where to find them online.
It also had the sequences of all the primers mentioned. The ones I tested were functional.
Some of the alleles (natMX6:P81nmt1-cls1-3xGFP:kanMX6 ; natMX6:Pnmt1-cls1-3xGFP:kanMX6 ; nem1Δ:NatMx6) were reproduceable, as proven in the .json files.
Assumptions
Found the Ase1 gene here: https://curation.pombase.org/allele_qc/genome_region?systematic_id=SPAPB1A10.09&format=genbank&upstream=0&downstream=0
Notes
fun experience! :)