What would be an ideal MOI for transducing neurons with AAV2 in culture around DIV 5? Any suggestions in improving the efficacy of transduction would be great.
I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...
03 March 2021 8,066 4 View
I have prepared a manuscript in IEEE Latex template, but for some purpose, I need to prepare a word file as well (detailed format is not required). Compared to two column .tex file, one column...
02 March 2021 1,830 3 View
Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue
02 March 2021 5,433 5 View
Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...
01 March 2021 2,027 2 View
I'm looking to try out an off-the-shelf EEG set for some exploratory work. Curious if anyone has used the OpenBCI, and how much set-up work is required. I've heard they are more difficult to get...
01 March 2021 4,128 3 View
I am doing a acid and a chiral amino alcohol reaction using DCC and HOBT in DCM or THF? Can anyone suggest me how to remove DCU and HOBT from a reaction mixture??
28 February 2021 2,153 1 View
Favourable conditions of base pressure, rate of deposition. Also, by which characterisation technique to be used to verify the thickness of the deposited film.
28 February 2021 3,640 3 View
The following code (see 1st 2 images attached) is used to produce PID controller values that are designed to control the system (G). The code finds the PID controller values (noted as k) by using...
28 February 2021 6,560 14 View
Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...
27 February 2021 5,028 2 View
What is the disadvantages of high sulfur content to the ceramic burning process? and what is the effect of outranged viscosity?
24 February 2021 8,219 2 View
Hi, I have been trying to transduce A549 and Calu-3 cells using the Lenti-X Tet-One inducible expression system (TakaraBio). My gene of interest is inserted in a pLVX-TetOne-Puro plasmid and has...
15 February 2021 8,462 3 View
I'm aiming to use an adoptive cell transfer approach, with microglia in mice with ALS. So far all the protocols I have seen, require sacrificing the mouse. I'd like to re-infuse the microglia back...
11 February 2021 9,509 1 View
It is required to evaluate shear modulus of concrete cube specimen of 100mm size. Since the diameter of DPC transducer is around 84mm, I would like to know whether it can give reliable shear wave...
02 February 2021 1,052 3 View
Hello, I hope to insert a gene into mammalian cells in culture using CRISPR/Cas9 and an HDR template. Will insertion of a 1 kilobase gene cause chromosomal abnormalities and effect cell...
31 January 2021 2,516 1 View
Hi, I'm trying to create a stable overexpression line with EF1a-FlagGOI-IRES-GFP using lentivirus packaged with pxPAX2 and pM2G. After transduction of 293T with 8ug/ml polybrene, I was able to...
29 January 2021 1,878 4 View
I have extracted the plasmid with the Qiagen maxi Endofree kit. the result of gel electrophoresis for this sample showed a band of degraded RNAs. Now, I want to remove RNAs from the eluted plasmid...
25 January 2021 2,958 3 View
I would like to troubleshoot the transfection stage of my lentiviral transduction protocol. In short, I have 4 lentiviral plasmids, one of which encodes GFP. I tried transfecting the HEK293T...
24 January 2021 2,341 3 View
I have been trying to express a gene (GFP) in a retroviral system by transfecting human Phoenix-ECO cells, and transducing murine PG13 cells with the supernatant. I then sort GFP-positive PG13...
18 January 2021 476 1 View
I want to generate a cell line expressing my gene under an inducible promoter. I chose FuW-tetO-MCS but I also need FUW-M2rtTA for the induction. So do I transfect both plasmids together in...
12 January 2021 8,501 2 View
I am trying to determine what best time-point to harvest my cells, HMVEC, after transduction with long non-coding RNA to determine changes in mRNA levels, specifically VEGFR2/KDR. I am capable of...
19 December 2020 4,865 2 View