PCR, Cloning, Restriction Digestion, Ligation, Transformation, Plasmid et al | Contact experts in Molecular Biology to get answers
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Questions related to Molecular Biology
Dear all, I used the EasySelect Pichia Expression Kit from Invitrogen with the pPICZα expression vector. The gene was cloned into PpiczαA vector at EcoRI and Xba1 site and was linearized with Sac1...
08 July 2019 3,218 6 View
Hello, I've isolated a bacteria from termite gut. After 16S ribosomal DNA sequence it was identified as Streptomyces sp. my strain was having cellobiohydrolase activity but i don't have it's...
18 May 2019 5,774 9 View
I want to fuse my gene of interest with EGFP and mCherry for localization studies. I am wondering if immdiate presence of mCherry or EGFT after my protein of interest would cause any steric...
21 March 2019 9,208 2 View
Hi all My DNA origami sample goes into a redsafe stained gel. I seperate the sample into species/bands containing the scaffold strand, the staples and the DNA origami. All bands appear...
04 March 2019 3,899 3 View
Actually I need a more clear explanation than does the Google.
23 February 2019 7,216 6 View
Thanks for your attention, I'd like to know whether all types endogenous RTases have a RNA binding domain or not. A research published on Nature Structural & Molecular Biology, declearred...
15 February 2019 7,167 4 View
I am trying to breed two types of Rosa26 knock-in mice together, but I have not had a single litter from 2 breeding pairs over 3 months. The breeders are now 6 months old. One breeder is a...
05 February 2019 7,747 2 View
I would like to detect a 6X-His tagged protein at the bacterial surface by immunofluorescence and FACS.
21 January 2019 5,183 4 View
I have extracted DNA from cacao leaf tissue (freeze dried). I used CTAB buffer (5%) along with 1% B-mercaptoethanol and 3% PVP. I did the DNA extraction overseas and transported it as a pellet to...
17 January 2019 2,152 12 View
I have extracted DNA from Cacao leaf tissue using CTAB, chloroform isoamyl alcohol method. When I quantified my samples on Nandodrop they show a conc. of 300-800 (ng/ul) in 50 ul of TE buffer. The...
16 January 2019 2,230 14 View
Promega is selling those Flexi-System Vectors. However, they come with Barnase as a selection marker and hence cannot simply be transformed into your usual library-competent E. coli strain. Since...
15 January 2019 650 4 View
I am trying to clone an 8 KB fragment to pFLAG-CTC vector. The vector map of the pFLAG-CTC vector is attached herewith. The issue I am facing is that after ligation reaction, and transformation...
12 January 2019 3,174 7 View
I have previously confirmed the presence of my (desired) plasmid after transforming it into bacteria and sequencing a colony. If I then transform that (purified) desired plasmid again into...
08 January 2019 7,304 8 View
Would you please suggest what can be reason if your control s 16 gene is not producing the expected results in bacterial sample in q PCR? While in same reaction the another gene showed absolutely...
28 December 2018 9,382 3 View
I was wandering is there any other way (instead of genetic ) such as using a chemical material or anything else to create thalassemia in the mouse?
16 December 2018 2,355 3 View
Hi there! I am trying to obtain a knock-out using Datsenko strategy. I am working with pKD4 (as a template for PCR) and the pKD46 vector. Once I obtained the linear DNA (using primers with 50 bp...
10 December 2018 2,813 10 View
Hi everyone, I am currently trying to clone a fragment (~1700bp) cut with ECORI into a vector (~5300 bp) linearized by ECORV. Since ECORI leaves cohesive ends I am using Klenow Fragment...
08 December 2018 10,055 5 View
I'm working on a presentation about history of protein complex research, connected to the paper about disordered protein complex by Borgia & Borgia in Nature. The idea is to show when new...
08 December 2018 1,308 4 View
I am searching for a distributor or collection of E. coli strains that offer classical strains for cloning (DH10b, DH5a, Rosetta, etc) to set up a reliable collection for cloning and classical...
03 December 2018 7,656 8 View
Hello, I'm interested in recent text books, review literature and possibly freely available online sources on classification, molecular biology and distribution of insect viruses. Any...
20 November 2018 3,308 2 View
I am trying to clone a promoter sequence using a pUC57 plasmid backbone in DH5alpha cells. The finished sequence will contain 6 direct repeats of a ~50 bp sequence I started from complimentary...
15 November 2018 373 6 View
Do you have an idea if there are molecular vectors available (other than viruses) that can be used in order to target specific genetic region and deactivate it specifically/in accurate way?. Like...
14 November 2018 7,522 6 View
I want to create a serial dilution of genomic DNA and plasmid samples for absolute qPCR. In order to get repeatable results in the Real time PCR, I want to do a very accurate dilution. I know that...
26 October 2018 5,388 12 View
Hi everyone, We are conducting gels to determine the structure of fungal community in soils. When we disclose the gels, the bands do not separate and they appear together in the middle or at the...
25 October 2018 8,349 6 View