Recently, I got a new lenti viral batch that has a titer of 9e6 +_3e6 TU/mL. The transduction efficiency that we got with the same MOI, that we used with the previous viral batch is much lower. Can anyone suggest me on this?
I have an input video, of skin (from a microscope that means it is magnified such that blood capillaries are visible). The idea is to extract 15 sec of continuous frames from a video of 1 min,...
02 March 2021 2,735 2 View
We have thin roots of from 1 year old different species of plants. We are looking into anatomical details of the roots. What's the best method to preserve and process the roots once they are...
23 February 2021 1,898 1 View
Hello We are working on a PCR product with an approximate size between 5000 and 7000 bp, we want to use it as a positive internal control, since we have sequenced it and it corresponds to the...
17 February 2021 2,896 3 View
I'm looking to do a sliding wear (plate-on-plate) simulation of GFRP composite plates with Archard Wear Model. Any hints or example steps are appreciated.
17 February 2021 1,447 2 View
There are numerous roots in this tap root system. Kindly suggest or share a research paper where I can clarify the doubts as to which root should be chosen for sectioning and for further root...
14 February 2021 1,816 3 View
We have a AAA class Newport solar simulator. We changed the Xenon Arc Lamp recently (XBO450W, Make:OSRAM) as the lifetime got over. However, the intensity is coming to be only 0.2 sun instead of 1...
14 February 2021 9,998 3 View
I am searching for tools to construct molecular beacons which shall be complementary to a given sequence. I don't want to design any primers. I found that UNAfold is one such tool which I could...
02 February 2021 9,065 5 View
I am synthesizing 7 amino acid long peptide sequence with CTC resin, 50oC coupling for 10 min. On cleavage (TFA/TIPS/Water 95:2.5:2.5) I got peptide with good yield. However, It has 68 Da more...
30 January 2021 8,483 3 View
Hello everyone, I am trying to prepare curvature map in ArcGIS 10.4, using spatial analyst tool. I am using 10 m CartoDem (projected in UTM 44N). I am getting very different value, curvature...
12 January 2021 504 4 View
I want to publish a case report regarding jejunal diverticula
02 January 2021 7,630 5 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
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 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
Hello, SV40 is not considered an oncogenic virus in this review:Oncogenic viruses and cancer Why is SV40 not considered an oncogenic virus when it produces the large T antigen, which is...
04 December 2020 8,374 5 View
Hello, I want to target a specific gene sequence using CRISPR-Cas9 to induce the death of the cells carrying that gene. I think that there are three basic approaches: 1.) Induce a frameshift...
20 November 2020 4,617 3 View