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Questions related to Sequencing
I am doing metagenomic sequencing without enrichment for an RNA virus. After my RNA is extracted from the blood I am doing library prep with the NEBNext Ultra directional RNA library prep kit for...
17 February 2021 5,234 2 View
I came across klenow fragment when I was reading about HiTS-FLIP method. I know klenow fragment doesn't posses 5'->3" exonuclease activity but what advantage does it provide for ?
02 November 2020 7,731 3 View
Hi everybody, I am doing CHIP experiment. The problem is that in my gel agarose I see alot of DNA with big size (longer that 1000 bp) which means there is alot of unfragmented DNA in my gel. I...
07 September 2020 1,612 5 View
Hi everyone, I have a question regarding sequencing SNPs. I have currently creating two SNPs on one gene and these SNPs are about 8kb apart from each other on two different exons. I would like to...
05 August 2020 5,666 3 View
I am interested in mRNAs only (also the ones from chloroplast). RNA input x RNA yielded approximately? Additional Kit used to concentrate RNAs + to get rid of other RNAs (tRNAs, etc...)? Thank...
02 June 2020 1,108 1 View
We have sequenced Synechococcus sp. PCC 7002 genome. Already the same strain has been sequenced and annotated by many before. Whole-genome map is also available online. The gene of interest is...
18 March 2020 1,061 12 View
Hello everyone! I am thinking to amplify T-DNA insertion region using LBb1.3 (left T-DNA border) and RP (gene specific Reverse Primer) against the genomic DNA of one of the T-DNA insertion mutant...
12 March 2020 1,353 3 View
The question is based on Sanger sequencing results. Gene size is 717bp which falls within the Sanger range. I was curious as to why did one of the reactions of the same sample show odd results...
26 February 2020 8,207 6 View
I am trying to figure out how many sequences I need for my samples. So far I can see that 40,000 would be necessary for microbial soil analysis. I have iDNA samples and so think I will need...
23 January 2020 896 1 View
Hi I have 20 RNA samples for miRNA sequencing because of its high cost, Some colleagues suggested pooling the samples, it means, splashing the samples into one or two sample and send for...
07 January 2020 5,179 3 View
Dear Researchers, I did Transcriptome of my two Fruit varieties at developmental, ripening and harvesting stage for the comparison of their differential gene expression profile, such as Sugar...
24 December 2019 6,637 4 View
Dear all, I cloned 25bp fragment into a vector. My no-insert ligation has no colonies and my insert+vector ligation has many. Which is the first clue that I have cloned the fragment. Than I...
23 December 2019 2,138 7 View
Dear All, 1 ug of permissible quality FFPE RNA is transcribed to cDNA through SSIV VILO RT kit. I have Qubit v.4 device, which can measure cDNA to a good extent. The cDNA ranges from 25-100 ng /...
12 November 2019 2,098 1 View
I've been doing some RF/Gibson cloning, and I am having issues sequencing my plasmids. I performed colony PCR to identify clones with the appropriate inserts. I then grew these up and plasmid...
22 October 2019 4,677 3 View
What are the criteria for designing biotinylated probes differ from primer designing?
25 September 2019 8,307 2 View
Hi. I amplified about 750bp of DNA from mushroom samples and got smearing bands in QC. what could be the reason? and what happens if I processed these samples for sequencing?
24 July 2019 3,249 6 View
I followed a regular CRISPR-cas9 transfection protocol, manually picked colonies into a 96-well plate and performed nested PCR which I then sent in for sequencing. In my previous 3 runs, I have...
12 July 2019 2,888 4 View
When we amplify 16S rDNA for sequencing using degenerate primers the amplified gene will result to amplicons with different sequences even with the same DNA band size. I assume that it will be...
28 March 2019 2,773 3 View
I am working with DNA methylation currently. I am creating hypomethylation condition in the cells. so how to confirm that the hypomethylation has occurred. sequencing of only LINE-1 gene will be...
20 March 2019 3,286 1 View
I've been using BioEdit for a decade but since it has no support and more like abandonware now I'd like to switch to a new sequence editing software which is being actively maintained. I would...
04 February 2019 7,382 6 View
I have mRNA sequence, from which i need to clip the UMI tag. Kindly suggest.
21 December 2018 1,126 2 View
Hi, everyone, I sequenced some samples in the Illumina MiSeq, got the results but now I don't know how to analyze them. I already read the User guide from Illumina, but it's only about the...
03 December 2018 9,489 3 View
If you were a researcher interested in investigating a particular group of prokaryotes (e.g. Cyanobacteria) from an environmental sequencing project, which of the following approaches to taxonomy...
27 October 2018 7,114 3 View
EDIT 29/10/2018: Added more details to bottom. This might be a bit weird, but I thought I'd try here anyway. Is it possible to configure Bowtie2, or similar programs, to match my query-sequence...
25 October 2018 7,448 2 View