eamag/protocols-with-steps huggingface dataset:
TITLE: Dendritic spine analysis
AUTHORS: Chuyu Chen

[DESCRIPTION]
Haloperidol is reported to induce a series of homeostatic synaptic and intrinsic adaptations primarily in the indirect pathway circuits. These adaptations include dendritic spine loss in iSPNs, which occurs after 5 days and persists for up to 14 days of daily treatment. We examined the haloperidol-induced dendritic spine density patterns in iSPNs after 7 days of treatment

[STEPS]
SECTION: Stereotaxic Surgeries
1. P4-5-day-old LRRK2GS/WT-Adora2aCre pups were cryoanesthetized and received ketoprofen for analgesia
SECTION: Stereotaxic Surgeries
2. Pups were placed on a cooling pad on a stereotaxic frame
SECTION: Stereotaxic Surgeries
3. 200 nl of the AAV-DIO-EGFP virus (7×10¹² vg/mL) were delivered into the dorsal striatum at a rate of 100 nl min-1 using Micro-4 controller (WPI).
SECTION: Stereotaxic Surgeries
4. After the injection, the pups were warmed on a heating pad before returning to their home cages.
SECTION: Confocal microscopy
5. Confocal images of fixed 80-um-thick brain sections of P30 pups injected with the AAV-DIO-EGFP were obtained with the Nikon A1R microscope
SECTION: Stereotaxic Surgeries
3.1. In order to ensure the dorsal striatum was targeted, the needle was placed 1 mm anterior to the midpoint between the ear and eye, 1.5 mm from the midline, and 1.8 mm ventral to the brain surface
SECTION: Confocal microscopy
6. Fluorescence projection images of dendrites and the corresponding dendritic spines were acquired with a 60x oil immersion objective (NA = 1.4) at 0.1 um intervals with 1,024 x 1,024 pixel resolution
SECTION: Dendritic spine analysis
7. Segments from secondary and tertiary dendrites without overlap with other neurons or discontinuities were chosen for analysis
SECTION: Dendritic spine analysis
7.1. Dendritic spine density was assessed using Imaris 10.1 software
SECTION: Dendritic spine analysis
8. Dendritic segments were traced using autopath mode of the filament tracer at default settings
SECTION: Dendritic spine analysis
9. Settings used for spine detection
SECTION: Dendritic spine analysis
9.1. Thinnest diameter of seed points at 0.45 μm; maximum length at 2.5 μm; no branched spines allowed; and seed point threshold at auto
TITLE: Glucose Tolerance Test
AUTHORS: Sabina Marciano, Roberta Marongiu

[DESCRIPTION]
Glucose tolerance test is performed to determine how quickly the glucose is cleared from the blood. It is used to test for diabetes or insulin resistance. 


[STEPS]
1. Single cage the mice for one week in advance
2. Weigh the mice
3. Fast the animals for 360 min 
4. After fasting, measure the glucose with a glucose meter
5. Perform an Intraperitoneal (IP) injection with 2 g/kg body weight of glucose (20% D-glucose stock solution dissolving 2g of glucose in 10ml saline and give 10ul per gram of body weight).
6.  Collect blood samples for glucose at 15, 30, 45, 60, 90 and 120 min post glucose injection
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
Both the iMicrobe (<a href="http://imicrobe.us" target="_blank">http://imicrobe.us</a>) and iVirus (<a href="http://ivirus.us" target="_blank">http://ivirus.us</a>) sites accept user data via an FTP (file transfer protocol) site (<a href="ftp://ftp.imicrobe.us" target="_blank">ftp://ftp.imicrobe.us</a>).  These are instructions for connecting and transferring data to us for inclusion in our database, the iPlant Data Store, and public availability via our FTP site.

[STEPS]
?. 
?. 
?.
TITLE: DRAGEN COVID Lineage App SARS-CoV-2 Strain Characterization on the Illumina BaseSpace Platform
AUTHORS: Technical  Outreach and Assistance for States Team

[DESCRIPTION]
This protocol provides instructions on how to run the DRAGON COVID Lineage app on the Illumina BaseSpace Sequence Hub. The DRAGON COVID Lineage app reads in fastq sequence files and produces Lineage, Clade, and Kmer Detection reports for each run. The information provided in these reports include Pangolin lineage classification, variant calls, and the fraction of human and SARS-CoV-2 kmers detected. The DRAGON COVID Lineage app also assembles a consensus sequence fasta file for each sample. This document applies to all whole-genome sequencing runs on the Illumina platform and downstream bioinformatics for public health laboratories.

For technical assistance, please contact: TOAST@cdc.gov

[STEPS]
SECTION: Create Basespace Project
1. Login to the Illumina BaseSpace Platform and create a new BaseSpace project
 

 

SECTION: Add Fastq files to BaseSpace Project
2. Add Fastq files to the new project directory. This can be done by either uploading fastq files from a local directory or by importing sequences using their Sequence Read Archive (SRA) accession numbers from NCBI using the ‘SRA Import’ app.   

 




SECTION: Run DRAGON COVID Lineage
3. Run the DRAGEN COVID Lineage App

 

 

 

SECTION: Access Summary Report
4. Accessing the DRAGEN COVID Lineage app summary report

 

 

SECTION: Downloading SARS-CoV-2 Consensus Sequences
5. Downloading consensus sequences generated by the DRAGEN COVID Lineage app

 


 

SECTION: Run DRAGEN COVID Lineage App
3. 
Run the DRAGEN COVID Lineage App

 

 

 

4. Accessing the DRAGEN COVID Lineage app summary report

 

 

5. Downloading consensus sequences generated by the DRAGEN COVID Lineage app

 


 


 

 

SECTION: Alternative Lineage Assignment
6. The SARS-CoV-2 consensus sequence assembly generated by the CDRAGEN COVID Lineage app can also be uploaded to other lineage assignment software.
SECTION: Alternative Lineage Assignment
6.1. Upload the consensus sequence for each sample to the Pangolin COVID-19 Lineage Assigner at:

https://pangolin.cog-uk.io/


Click the 'Start analysis' button:

  

The Pangolin COVID-19 Lineage Assigner returns the lineage classification and assignment probability:

  

SECTION: Alternative Lineage Assignment
6.2. Or upload the consensus sequence for each sample to the Nextclade clade assignment web portal at: 

https://clades.nextstrain.org/

  

The Nextclade server provides clade classification as well as QC metrics and a list of amino acid substitutions. A summary output file can be downloaded with the 'Export to CSV' button.
TITLE: Adding 1-hydroxyphenazine to L4 C. elegans in liquid culture
AUTHORS: Muhammad Zaka Asif, Man Shah, Yosef Smadi

[DESCRIPTION]
This protocol describes day 4 of our workflow to grow worms in liquid culture to induce the production of natural products (in this case, 1-HP derivatives). In this protocol, we add the toxin to our worms grown in liquid culture to induce the production of glycosylated products. 

[STEPS]
1. Follow steps 1-6 from the protocol for transferring C. elegans to S-buffer, but centrifuge at 525 RCF for 1 minute
2. Dilute to a final concentration of 30,000 worms/mL in S-basal and final toxin concentration of 22.3 µM 1-HP. 
3. Repeat steps 1-2 for 1.1% DMSO and bacteria-only controls.
4. If using negative controls, repeat steps 1-3 for them.
5. Put the flasks back in the shaker and allow to incubate for 24 hours.
TITLE: easyDB – Circularization of rv0678 for genotypic bedaquiline resistance testing of Mycobacterium tuberculosis
AUTHORS: Jason D Limberis

[DESCRIPTION]
We designed primers with a tail sequence that forms a six-nucleotide hairpin at temperature <55oC, but not ≥55oC. These primers contain six phosphorothioate bonds starting at the complementary region to inhibit exonuclease T7 activity. The primers successfully amplified the target and, following incubation with a mixture of T7 exonuclease, DNA polymerase, and Taq DNA ligase, pseudo-circular double-stranded DNA formed.

[STEPS]
SECTION: Prepare Buffers
1. 

      ISO buffer   (2.5X)  Volume (ul)    1M Tris-HCl pH 7.5  100    200mM MgCl2  50    100mM dGTP  2    100mM dATP  2    100mM dTTP  2    100mM dCTP  2    100mM DTT  100    PEG 8000  50mg    50 mM NAD  20    H20  90    
 
      Master Mix  Volume (ul)    2.5X ISO buffer  640    T7 exonuclease (10 U/μl)  0.64    2 U/μl Phusion polymerase  20    40 U/μl Taq DNA ligase  160    H20  379.36  
 
SECTION: Amplicon PCR
2.  
      Component  Volume (ul)    5X Reaction Buffer  10    5X Q5 High GC Enhancer  10    10 mM dNTPs  1    easy_HP_v2_Rv0678_lng_v2_F  2.5    easy_HP_v2_Rv0678_lng_v2_R  2.5    DNA (5ng)  2    Q5 High-Fidelity DNA Polymerase  0.5    Nuclease-Free Water  21.5  
 
 
 

SECTION: Amplicon PCR
3. Add 40 µL of resuspended AMPure XP beads
Mix by pipetting 10x
Incubate 5 min at 65 Room temperature
Place on magnet
Wash 2x with 200 µL freshly-prepared 70 % (v/v) ethanol
Air dry for30 s, don't allow the beads to become cracked
Resuspend in 20 µL Tris-low EDTA
Mix by pipetting 10x
Incubate 5 min at 65 Room temperature
Place on the magnet, aspirate  20 µL of the eluant into a new 200ul tube
SECTION: Exonuclease Treatment – optional
6. Optional
 
      Component  20ul reaction    H20  7    Cutsmart  2    DNA  10    Exonuclease   VIII, truncated  0.5    Exonuclease III  0.5  
 
SECTION: Exonuclease Treatment – optional
7. Add 20 µL of resuspended AMPure XP beads
Mix by pipetting 10x
Incubate 5 min at 65 Room temperature
Place on magnet
Wash 2x with 200 µL freshly-prepared 70 % (v/v) ethanol
Air dry for30 s, don't allow the beads to become cracked
Resuspend in 12 µL Tris-low EDTA
Mix by pipetting 10x
Incubate 5 min at 65 Room temperature
Place on the magnet, aspirate  20 µL of the eluant into a new 200ul tube
SECTION: easyDB reaction
4. Thaw a 10ul aliquot of easyDB Master Mix on ice
Add 5ul (~150ng) DNA to the tube
Mix thoroughly by pipetting 10X
Incubate at 50°C for 60min (will be reduced, probably to 10min)
 
SECTION: easyDB reaction
5. Add 20 µL of resuspended AMPure XP beads
Mix by pipetting 10x
Incubate 5 min at 65 Room temperature
Place on magnet
Wash 2x with 200 µL freshly-prepared 70 % (v/v) ethanol
Air dry for30 s, don't allow the beads to become cracked
Resuspend in 12 µL Tris-low EDTA
Mix by pipetting 10x
Incubate 5 min at 65 Room temperature
Place on the magnet, aspirate  12 µL of the eluant into a new 200ul tube
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This is the hands-on interactive component to the ECOGEO Workshop Module on Binning.</p>

[BEFORE_START]
<p>This tutorial is tailored for <a href="http://anvio.org" target="_blank">Anvi’o</a> v2.0.1. If you do not have this version available to you, certain things may not work the way they should. The purpose of this tutorial is to demonstrate some of the anvi’o capabilities using the Infant Gut dataset, which was generated, analyzed, and published by <a href="http://www.ncbi.nlm.nih.gov/pubmed/22936250" target="_blank">Sharon et al. (2013)</a>, and was re-analyzed in the <a href="https://peerj.com/articles/1319/" target="_blank">anvi’o methods paper</a>.</p>
<p> </p>
<p>The attached ecogeo-binning-tutorial.pdf is the full tutorial used during the actual workshop. Many of the links no longer work and should be followed from this protocol. A similar non-ECOGEO tutorial is also updated and available online (<a href="http://merenlab.org/tutorials/infant-gut/" target="_blank">http://merenlab.org/tutorials/infant-gut/</a>)</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Marchantia spores production in Microboxes
AUTHORS: Eftychis Frangedakis

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Spore sterilisation</div><div class = "text-block"><span>Marchantia spores can be obtained usually in three months using Microboxes. The lid of Microboxes has a specially designed filter that allows high gas exchange, limited dehydration and blocks the entry of contaminants such as fungi into the culture. We are using the TP4000+TPD4000 Microboxes with</span><span style = "font-weight:bold;"> </span><span style = "font-weight:bold;font-weight:bold;">a</span><span style = "font-weight:bold;"> </span><span>green XXL+ filter. </span></div></div>

[STEPS]
?. Put 15 “Jiffy 7” pellets in a Microbox, add 800 mL of water, close the lid and autoclave (A and B in Figure).
?. After autoclaving the Microbox with the “Jiffy 7” pellets, work in a flow hood.
?. Put at least one 10 mm x 10 mm thallus fragment or 3 gemmae per pellet using sterile tweezers (C and D in Figure).
?. Add another 150-200 mL of sterile water, close the lid and place at 21 °C under continuous light with light intensity of 150 μmol/m2/s (E in Figure).
?. After one month transfer plants under a 16 h light / 8 h dark regime, with light intensity of 150 μmol/m2/s supplemented with far red light (peak emission: 700-750 nm, spectral photon flux density: 2400 μmol/m2/s, distance from plants: 32 cm) (F in Figure).
?. After approximately 4 weeks mature male and female reproductive organs are produced (G and H in Figure).
?. In a flow hood, open the lid of the Microbox, and using a pipette add 5-10 mL of sterile water on top of the male and female reproductive organ (I and J in Figure). After a few seconds you will be able to see the sperm as a cloudy exudate in the water drop. For fertilisation, use the pipette to "move" the reproductive organs in order to touch each other.
?. Alternatively, for fertilisation, transfer the water drop from the male gametophore with a disposable sterile plastic pipette on the top of the female reproductive organ (pic in step 11).
?. After one 4 weeks mature sporangia are ready for collection (K in Figure). Place 1-3 mature sporophytes in a 1.5 mL Eppendorf tube with 5 silica gel beads (L in Figure) and store at 4 °C if the spores are to be used within 1-2 months. For longer storage, allow the sporophyte to desiccate for a week at 4 °C and then store at -80 °C.
?. 
?.
TITLE: Peptide Drug Screening
AUTHORS: Cathy  Miller

[DESCRIPTION]
Peptides are ideally suited to mimic natural ligands and usually act in an antagonistic or agonistic manner. Small active peptide molecules have huge potential to be developed as vaccines, diagnostic reagentsand lead compounds. Therefore, peptides have become the highlight of drug discovery, especially due to their small size, structural diversity and high specificity for a given target. Moreover, peptides can be selected from large natural or artificial peptide libraries rapidly, and are easy for chemical processing and synthesis.

Application
Peptides are capable of physiologically destroying functional complexes due to their small size and specific binding properties.
Peptides perform a variety of biological functions in the body, such as metabolism, digestion, respiration, reproduction, immune defense and many other functions by acting as ligands (such as hormones, neurotransmitters, cytokines, and chemokines).

Our Peptide Drug Screening Services
Creative Peptides provides a wide range of screening strategies to meet the needs of every customer for peptide drug screening on time and budget. Our skilled and dedicated scientific researchers have rich experiences in selecting the most suitable methods and techniques for each peptide drug project. Over the years, the excellent quality and reliability of our peptide services have been appreciated by customers worldwide.

[STEPS]
TITLE: 70% Ethanol Fixation
AUTHORS: Arnold  Federico

[DESCRIPTION]
This protocol is specifically written for fixing/permeabilizing suspension cell culture in a final volume of 1mL. For DNA content analysis, such as for cell cycle analysis, a precipitating fixative such as ethanol is preferred over formaldehyde fixation (Darzynkiewicz et al. 2001). The process of fixation preserves the cell state, makes cells permeable and allows for storage for extended periods of time.

[STEPS]
SECTION: Protocol
1. Transfer 1 * 106 to 5 * 106 suspended cells to appropriate centrifuge tube and pellet by spinning at 300g for 5 minutes

SECTION: Protocol
2. Discard supernatant without disturbing pellet. Resuspend and wash cells in 1mL 1X PBS with gentle pipetting. Spin down cells again at 300g for 5 minutes. 
SECTION: Protocol
2.1. While cells are spinning, add 700uL of 100% EtOH (pre-cooled in -20ºC)  to new 1.5mL microcentrifuge tube and keep on ice
SECTION: Protocol
3. Discard supernatant without disturbing pellet. Resuspend pellet in 300uL 1X PBS and gently pipette or vortex to break up cell clumps. (Cell aggregates will be stabilized when introduced to ethanol, make sure you have an even cell suspension!).

Transfer all 300uL cell suspension to microcentrifuge tube containing 700uL EtOH on ice and pipette mix to evenly distribute cells.
SECTION: Protocol
4. Transfer cells to -20ºC  for at least 2hr prior to staining/analysis. Cells should be stable in -20ºC for months.
TITLE: HannerLab Qubit Protocol
AUTHORS: klindsay

[DESCRIPTION]
The Qubit can measure the concentration of DNA (in ng/µL).


•  Qubit HS (High sensitivity) is used for samples with low traces (0.2-100ng) of DNA (ie. eDNA)


• Qubit BR (Broad range) is used for samples with higher traces of DNA (2-1000ng) (ie. Tissue)


•The Qubit uses specially-sized tubes for readings (see materials)
Tip: These tubes fit perfectly in an empty 1000ul tip box, or on the bottom of most tube racks.

Make sure to label your Qubit tubes so you know which sample they correspond to when taking the readings. 




[STEPS]
1. Prepare Qubit Buffer and Dye mix for the number of samples + 2 (for standards) + 1 extra.
1.2. •	Solution will be 199 parts buffer and 1 part dye (e.g., 199uL buffer for every 1uL dye)
1.1. •	(# of samples + 3 volumes)
2. Vortex buffer-dye mix, then aliquot 198uL buffer-dye mix into each Qubit® tube for each of your samples
3. Aliquot 190uL buffer-dye mix into each Qubit® tube for standard 1 & 2
4. Pipette 10uL of each standard into its designated Qubit® tube (Standard 1 & 2)
4.1. •	The standards are stored in the black fridge near the back of the lab
4.2. •	Mix solution with pipette
5. Pipette 2uL of each sample into its designated Qubit® tube
5.1. •	For detection with low-yield DNA, mix 197ul buffer-dye with 3ul sample instead
5.2. •	Mix solution by pipetting up and down up to 10 times.
6. Turn on the Qubit® and select "dsDNA High Sensitivity" or "brDNA Broad range", depending on which kit you used
7. Insert Standard 1, close the hatch, and Click "read standard"
8. Repeat for Standard 2
9. Select “2uL” when asked about sample volume (or 3ul if you used 3) as well as “ng/uL” for concentration units
10. Insert the first sample tube, close the hatch, and select “Read Sample”.
11. Record concentration
12. Repeat for all samples
13. Re-measure all samples two more times and take an average of the three values for each sample. Wait 3-10 minutes between each reading. 

NOTE: If you are getting an "out of range reading" (ie. too low) make sure to shake the tube downwards so that no liquid is trapped at the lid. Consider using a different kit if you are consistently getting out of range values (ie. too low means you should be using the HS kit, too high means you should be using the BR kit). It is normal for values to fluctuate slightly as time progresses as the dye as more time to interact with the DNA in the sample.
11.1. Sometimes by pressing the "read tube" button for too long, you may cancel the reading. Make sure the new reading is not identical to the last. If unsure, read the tube again. 
1.3. 

e.g. for 10 samples the calculation would be (10 + 2 + 1)

So your buffer-dye mix will be:
199uL x 13 = 2587uL mixed with 
1uL dye x 13 =13uL

For a total of 2600ul. Make sure to choose the appropriate tube volume (2ml, 5ml, 20ml) for your buffer-dye mix
TITLE: GST fusion protein production
AUTHORS: Leonardo A Parra-Rivas

[DESCRIPTION]
GST fusion protein production

[STEPS]
1. Full-length recombinant human WT α-syn, α-syn S129A, and S129D were expressed in Escherichia coli BL21 (DE3) (New England Biolabs, Cat # C2530H) using the bacterial expression vector pGEX-KGmyc. Following transformation protein expression was induced with 0.05 mM IPTG (isopropyl-β-d-thiogalactopyranoside),and either incubated at 37 °C for 2 hours or at room temperature for 6 hours,
with shaking. 






2. The cells grown on Terrific Broth (Thermo Scientific Cat#
BP9728-2) were harvested by centrifugation at 4500× g at 4 °C for 20 min, and pellets were stored
at -80 °C until use. 
3. For protein purification, protein pellets were resuspended
in 30 ml Lysis Buffer containing 1X PBS, 0.5 mg/ml lysozyme, 1 mM PMSF, DNase,
and EDTA-free protease cocktail inhibitor (Roche Cat# 11836170001) for 15 min
on ice, briefly sonicated (3 sets with 33 strikes and 30-second breaks on ice
between sets), and removed the insoluble material by centrifugation at 15,000 g
at 4 °C for 30 min. 
4. The clarified lysate was incubated with 500 ml of glutathione-Sepharose
4B (Sigma Cat# 17-0756-01), preequilibrated with 1X PBS containing 0.1% Tween
20 and 5% glycerol (binding buffer), on a tumbler at 4 °C overnight. 
5. The GST-bound proteins were washed four times with 30 ml binding buffer and
maintained at 4 °C for pull-down assays, in vitro phosphorylation, and
in vitro dephosphorylation experiments. 
6. GST-bound proteins were occasionally eluted by TEV cleavage. In brief, 15 ug of the fusion protein was mixed with 5 ul of TEV protease reaction buffer (10X) and 1ul of TEV protease
(New England Biolabs, Cat#P8112), followed by overnight incubation at 4 °C.
TITLE: Tissue Sectioning Guidelines (Fresh Frozen)- CODEX/Phenocycler-Fusion
AUTHORS: Santhosh Sivajothi

[DESCRIPTION]
The purpose of this SOP is to outline the techniques for sectioning and storage of fresh-frozen tissue samples for PhenoCycler-Fusion experiments

[GUIDELINES]
Fresh-frozen tissue sections are mounted directly onto slides. Appropriate preparation and storage
of tissue sections are critical to ensure sample integrity.

Tissue sections adhered directly onto slides can be stored at -80°C for up to 6 months before
staining.
Tissue thickness must not exceed 10 μm as this can affect the autofocusing capabilities of the
microscope.
For best results, tissue sections should be completely adhered to the slide without folds or tears.
To ensure that tissue sections are not damaged, it is critical that the tissue slides are not
stacked on top of one another

[STEPS]
SECTION: PREPARE CRYOSTAT CHAMBER
1. Standard cryostats with temperature control are recommended for tissue sectioning. Most tissues are sectioned in temperatures ranging from -15°C to -25°C. The exact temperature is unique to each tissue type and should be determined according to standard sectioning procedures.
SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
2. Set the cryostat chamber to tissue-specific temperature range.
SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
3. Place the slide storage box in the cryostat chamber to equilibrate to the cryostat temperature.
SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
4. Once the cryostat reaches the programmed temperature, transfer the tissue from the - 80°C freezer to the cryostat using a container filled with dry ice.
SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
5. Use compressed air to remove dust and lint from the slides before use.
SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
6. Place the prepared slides in the cryostat chamber to equilibrate temperature for 20-30 seconds.
SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
7. Section the tissue at a thickness of 5-10 μm.
 


SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
8. Gently place the tissue section in the center of the slide within the imageable area as shown in
grey rectangle below. Multiple tissues can be placed within this region as long as there is no overlap of tissue-on-tissue or OCT-on-tissue.


  
 

SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
9. Adhere the tissue section to the slide by placing a gloved finger underneath the slide for 1- 2
seconds
 

SECTION: FRESH-FROZEN TISSUES - SECTIONING PROCEDURE
10. Place the mounted slide in a single slot of a microscope slide box. Once complete, cover the slide storage box with the lid. Place the box of mounted slides on dry ice for transport to a -80°C freezer. 

Store slides at -80°C for up to 6 months