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Questions related to Cloning
I am trying to stain stage 17 embryos using phalloidin without any success. I am fixing the embryos in a 1:1 mixture of heptane/4% PFA after dechorionation in 50% bleach. I devitellinize by hand...
30 January 2022 2,256 1 View
I have a CRO performing mouse assays but they don't have in house molecular biology facility and we have to transport spleens to a different facility in a different city. We can expedite shipping...
25 January 2022 5,475 2 View
I want to check if I can use GFP mice for IF quantification and look for proteins using secondaries that wouldn't excite GFP? (568 or 532)? Would GFP signal interfere with my staining and...
05 January 2022 5,172 2 View
I am working on a project where I am designing an aerosol formulation of my therapeutical peptide. I would like to do the study of biodistribution in vivo (mice), so I used cy5-labelled peptide...
20 December 2021 431 1 View
Hello, I would like to include a GFP tag in the plasmids I am preparing. I found multiple sequences for GFP and I would like to understand the differences between them. Does anybody have a...
19 December 2021 6,483 1 View
So I have always used Hygromycin/Kanamycin selection for Arabidopsis but this is my first time I am using methotrexate for selection of T0 transformed plants. Plants with hygromycin resistance I...
13 December 2021 8,121 1 View
Hi all, We're trying to re-capitulate some of the amazing papers showing excellent production of IgG in Sf9 and other insect cell cultures. However, we've been completely unable to find any...
09 December 2021 1,291 1 View
Hello, i was trying to alignning multiple sequence with MEGA for constructing phylogenetic tree. after aligning, I am supposed to trimming sequence to make them share equal length. But i...
07 December 2021 3,329 3 View
Hello, I am relatively new to using electroporation transformation. Recently I cam across a problem transforming two plasmids (one with >10 kbp and the other approx. 6 kbp) into MG delta lacI...
23 November 2021 7,139 8 View
I have a question, I try to isolate phages from environmental samples, and during this process, I face with a clear glass tubes after overnight incubation (host+phage), but I spot nothing on the...
18 November 2021 437 6 View
I am using FFPE tumor samples that are used for laser microdissection (Leica). Prior to laser microdissection, the tissue is deparaffinized and stained with hemotoxylin. LMD samples are stored in...
11 November 2021 9,889 0 View
Hi RNA experts, I am trying to extract RNA from my blueberry fruit samples. They have been stored in -20 C for one or two months and then they were moved to -80 C. I used the Qiagen RNeasy Plant...
11 November 2021 144 6 View
Would it harm if an MDD patient receives TMS sessions 6 days a week instead of 5? On a different note, is it required for the patient to receive 5 sessions in a row, or a patient could have TMS...
04 November 2021 4,661 4 View
How conformational changes are occurring in protein denaturation related to the Gibbs free energy changes of that particular protein? For example I have one protein having two domain in its native...
01 November 2021 5,704 2 View
We were comparing the diversity of moist and dry broadleaf forests as part of our Ecology project. It was found that Shannon diversity index is greater for moist broadleaf forest and Simpson...
31 October 2021 3,203 5 View
I have simulated in silico how to clone a large protein (1900bp) Please, any suggestions on what measures to consider before doing wet lab to clone and express this protein? Thank you for your...
29 October 2021 2,803 5 View
I´ve seen some toxin-antitoxin systems in bacterial chromosomal regions that are not conserved in related genomes. In addittion to their role in plasmid manteinance, are these systems reported to...
28 October 2021 9,032 1 View
I don't have KH2PO4 but I have KCL.
19 October 2021 6,426 3 View
Hello, I'm relatively new to the field of Phage Display, and there are still points I don't perfectly understand. A bit of context : I'm using M13 phage along E.Coli for amplification/infection....
15 October 2021 2,924 3 View
I want to silence a gene with a medical purpose in mind. From what I read, the use of shRNA is frowned upon due to toxicity concerns, with researchers turning instead to artificial miRNA for the...
29 September 2021 5,425 0 View
I'm currently trying to delete a gene from a Gram-negative environmental bacteria by using CRISPR/Cas9. However, I can't find good literature or protocol for introducing Cas9/SgRNA complex to a...
28 September 2021 3,951 5 View
I am working with various strains of E. coli for protein expression by cloning. Recently, I observed that the colonies of E. coli C43(DE3) is different from C41(DE3). The C41(DE3) colonies are...
22 September 2021 5,822 3 View
Primers Oligo synthesis purification PCR
20 September 2021 8,379 3 View
My question concerns the import of proteins into the endoplasmic reticulum and the nucleus.
10 September 2021 5,016 1 View