Which step is most crucial for DNA isloation???

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RNA extraction from pancreas?

We tried RNA isolation from murine pancreas many times but we are getting degraded RNA. Kindly suggest precaution for the same.

11 December 2017 7,489 7 View

Does SNP in 3' UTR region affects over all protein function?

SNP in the 3' UTR region is involve in any kind of mechanism. These SNPs play any role in protein function, if yes how? How can one find that?

04 May 2017 2,369 6 View

What is the most crucial step for RNA isolation from human adipose tissue?

I am trying to isolate RNA from human adipose tissue but I am not able to get it. I have already tried from PBMCs and other cell type, from which I am able to isolate.

01 February 2016 7,068 4 View

Why am I getting lot of non-specific pcr band?

I have already tried decreasing primer concentration, Touch-down PCR and increasing annealing temperature. My pcr product is of 200bp. Gel image of my pcr product with two different dna samples,...

31 December 2015 9,789 8 View

Can someone help me find the PENHAPLO program for haplotype & phenotype analysis?

PENHAPLO program for haplotype & phenotype analysis

01 January 1970 1,205 3 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Does crude extraction using NaOH and Tris work well with Fungi?

I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you

08 August 2024 4,733 2 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Do you have good tips for seaweed tissue preservation in the field for post RNA extraction?

I will be with my students collecting seaweed samples in a marine farm and later we will process this tissue for RNA isolation and further sequencing. Does anyone have tips on how to collect the...

04 August 2024 501 2 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,420 1 View

Is it common for heterotrophic bacteria to grow on Pro99 and SNAX media?

I am attempting to isolate picocyanobacteria from seawater (pre-filtered with a GF/D membrane) using the pour plate technique and filter plating method (Kearney et al., 2022) on Pro99 and SNAX...

30 July 2024 1,040 1 View

Are the apoptotic cells is positive for γH2AX ?

It has been documented that apoptotic cells themselves can induce phosphorylation of serine 139 on H2AX (γH2AX) due to DNA fragmentation during apoptosis (doi: 10.1074/jbc.275.13.9390). As γH2AX...

28 July 2024 7,983 2 View