METADATA
last updated: 2026-02-16 BA after RT initial conversion
file_name: SOP-301 PGS48 Verification - Run Form.md
file_date: 2022-01-01
title: FloodLAMP QMS SOP-301 PGS48 Verification - Run Form
category: guides
subcategory: manufacturing
tags: 
source_file_type: gdoc
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gfile_url: https://docs.google.com/document/d/1mVuPCxzU2bUILebEfg7gvHwjzD6DwGqv
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pdf_gdrive_url: https://drive.google.com/file/d/1W7vkFU0raZso77GsFPm3LWNDMkqKmwm1
pdf_github_url: https://github.com/FocusOnFoundationsNonprofit/floodlamp-archive/blob/main/guides/manufacturing/SOP-301%20PGS48%20Verification%20-%20Run%20Form.pdf
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conversion: pandoc
license: CC BY 4.0 - https://creativecommons.org/licenses/by/4.0/
tokens: 1198
words: 671
notes: 
summary_short: The FloodLAMP QMS SOP-301 is a verification run form for testing PGS48 (Primer-Guanidine Solution) batches via CLAMP amplification. It covers preparation of reaction mixes (46 µL PGS + 55 µL CLAMP MM per tube), positive/negative control setup, and a plate layout map for tracking POS/NEG results by PGS lot number. The second half includes an Amp Run Sheet Short with standard amplification procedures, heater settings (65°C for 25 min), and write-in fields for operator documentation and result logging.


CONTENT

FloodLAMP Biotechnologies
**Document Number:** SOP-301-A
**Effective Date:** 01/XX/2022
**Version:** 1

***INTERNAL TITLE:*** PGS48 Verification - Run Form

## PREP
- Amp heaters on (Heat indicator lit!)
- Map out plate layout
- Printout these 2 pages out separate
- Label Reaction Mix tubes
  - 1.5mL EDLB tubes
  - 4 rxn per RM tube
  - 1 RM tubes per PGS batch (#68)
- Calc amount of CLAMP MM needed
  - 55 µL x # batches = \_\_\_\_\_\_\_\_\_
  - Use single source tube
  - If larger run combine MM tubes
- Label strip8 tubes (or use plate)
- Make 1mL 1XISS (fill out on next pg)

## PREP USER WRITE IN
_QR Code_ Validation PGS
[Form Link](https://docs.google.com/forms/d/e/1FAIpQLSdGHu4xGNY6SDBQqnp0ZH3N4lL0r4EnNr7e3cYC8mKiNyIXag/closedform)
Location: MBC
Name:
Date and Time:

## RUN
- Get PCR cold block out 10min before - put plate or strip8 tubes in
- Get blue tube cold block out 10min before - keep all RM tubes in this
- Make Reaction Mixes for all tubes:
1. Add 46 µL of each PGS to each RM tube (making sure to match lot#'s)
2. Add 55 µL of CLAMP MM to each RM Tube
3. Vortex for 10s and centrifuge/shake down
4. Add 23µL of reaction mix to each of 4 wells {at bottom, no blowout, no tip touch}
- After adding RM to all reconfigure POS strips together and NEG strips together
- Add NEG control (1XISS) to those columns/tubes - cap
- Prep Pos Control: TPC100 - thaw 30s, add 20uL of Ultra Pure water (clean glove!)
- Add POS - TPC50 to each POS tube PGS Lot# Prefix \_\_\_\_

|  | POS | NEG | POS | NEG | POS | NEG | POS | NEG | POS | NEG | POS | NEG |
|------|------|------|------|------|------|------|------|------|------|------|------|------|
|  | **1** | **2** | **3** | **4** | **5** | **6** | **7** | **8** | **9** | **10** | **11** | **12** |
| **A 1** | POS 0040 | NEG 0040 |  |  |  |  |  |  |  |  |  |  |
| **B 2** | POS 0040 | NEG 0040 |  |  |  |  |  |  |  |  |  |  |
| **C 3** | POS 0041 | NEG 0041 |  |  |  |  |  |  |  |  |  |  |
| **D 4** | POS 0041 | NEG 0041 |  |  |  |  |  |  |  |  |  |  |
| **E 5** | POS 0042 | NEG 0042 |  |  |  |  |  |  |  |  |  |  |
| **F 6** | POS 0042 | NEG 0042 |  |  |  |  |  |  |  |  |  |  |
| **G 7** |  |  |  |  |  |  |  |  |  |  |  |  |
| **H 8** |  |  |  |  |  |  |  |  |  |  |  |  |
||

## PREP
- Heaters on (Heat indicator lit!)
- 1X Inactivation Saline Soln ready
- Reaction mix strip8/plates ready
- Safety Procedures:
  - lab coat
  - gloves
  - face mask
  - face shield or goggles
- Alcohol wipe sample tubes

## PREP USER WRITE IN
|               | Amp Run Sheet Short Any System | version 1.3mod |
|---------------|-------------------------------|----------------|
|               |                               |                |
|               |                               |                |
|               |                               |                |
||

## AMPLIFICATION REACTION
- Thaw Reaction Mix (if frozen)
- Setup tips to align with reactions
- Add 2μL from each sample tube (pipet up & down 5X, blowout in liquid, tip touch)
- Heat 65°C for 25 min, set timer
- Intake sample tubes in App
- Put sample tubes in fridge
- Remove Reactions and Let cool for 1 min before photo
- Take photo in lightbox
- Crop photo and apply vivid filter
- Update results in App
- Log Run with Form link

## AMPLIFICATION REACTION USER WRITE IN
Amp Heater: Strip8 Tubes or Plates
Reaction Mix: Fresh
CLAMP MM Lot and EXP:
Num Reactions (including controls):
Pos Ctrl ID: TPC50 from TPC\_
Ultrapure Water ID:
Neg Ctrl ID (today's 1XISS):
Initial Inconclusives?:
Notes: