I use T4 ligase buffer for Golden Gate because it doesn't contain PEG. However, I don't know its compatibility with BpiI. I was wondering if anyone has experience with this?
Hello. Recently I've been assembling plasmids that use pSMPUW as the backbone (kanamycin resistant). I've noticed that the minipreps for these plasmids usually sequence fine (I've been using stbl3...
03 April 2019 6,600 2 View
I have a vector containing the EF1a promoter. I was wondering if DH5a cells will accept this vector upon transformation? I know some bacteria are incompatible with certain gene segments.
02 March 2018 2,336 8 View
Hello. So for a current project I'm doing, I'm assembling several fragments (6-7) using golden gate assembly. I've combined the ligation and digestion in to one reaction, which takes ~5 hours in...
08 September 2017 1,762 2 View
Say you have three fragments that have overlaps with each other. During PCR, I'm assuming that since these fragments could physically overlap with each other they will anneal. My question is will...
07 August 2015 2,152 4 View
Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?
07 August 2024 4,616 0 View
Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...
06 August 2024 5,373 2 View
Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...
05 August 2024 3,783 2 View
Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC
01 August 2024 967 3 View
Given that the bacterial genome has over 800 contigs, but its quality metrics are good, with a completeness of 98.55% and a contamination of 0.68% as assessed by CheckM, what specific validation...
01 August 2024 1,514 1 View
Hello everyone! How can i prepare EDTA buffer solution (pH 8.2) to determine GSH levels? Thank you in advance.
29 July 2024 4,374 1 View
I want to preserve the sample leaves for the LM study of the stomata and conidia analysis. Whether 2.5% glutaraldehyde in 0.05 M phosphate buffer with pH 7 can be applied for preservation.
29 July 2024 8,560 0 View
// interested in the difference between floating events and short circuits.
22 July 2024 6,565 0 View
Our lab has extracted bacterial DNA samples using Qiagen DNeasy kit and eluted the samples in AE buffer. However, this buffer contains 0.5mM EDTA not suitable for downstream application. Any ideas...
13 July 2024 1,675 3 View
Commonly, we prepare 1.0% of agarose gel with 0.4g agarose powder and 40ml 1X TAE buffer. if I would like to prepare a 1.2%% of agarose gel, is it I need to add 0.48g agarose powder and 120ml 1X...
12 July 2024 2,058 5 View